Glucoamylase variants with altered properties

ABSTRACT

The present disclosure relates to variants of a parent glucoamylase having altered properties (e.g., improved thermostability and/or specific activity). In particular, the present disclosure provides compositions comprising the variant glucoamylases, including starch hydrolyzing compositions and cleaning compositions. The disclosure also relates to DNA constructs encoding the variants and methods of producing the glucoamylase variants in host cells.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims benefit to U.S. Provisional Application No.60/989,426 filed Nov. 20, 2007.

SEQUENCE LISTING

Also attached is a sequence listing comprising SEQ ID NOs: 1-167, whichis herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

Glucoamylase variants advantageously have altered properties (e.g.,improved thermostability and/or specific activity). Compositionscomprising the variant glucoamylases, DNA constructs encoding thevariants, and methods of producing the glucoamylase variants in hostcells are provided.

BACKGROUND

Glucoamylase enzymes (glucan 1,4-α-glucohydrolases, EC 3.2.1.3) arestarch hydrolyzing exo-acting carbohydrases, which catalyze the removalof successive glucose units from the non-reducing ends of starch orrelated oligo and polysaccharide molecules. Glucoamylases can hydrolyzeboth the linear and branched glucosidic linkages of starch (e.g.,amylose and amylopectin).

Glucoamylases are produced by numerous strains of bacteria, fungi, yeastand plants. Particularly interesting, and commercially important,glucoamylases are fungal enzymes that are extracellularly produced, forexample from strains of Aspergillus (Svensson et al. (1983) CarlsbergRes. Commun. 48:529-544; Boel et al., (1984) EMBO J. 3:1097-1102;Hayashida et al., (1989) Agric. Biol. Chem. 53:923-929; U.S. Pat. No.5,024,941; U.S. Pat. No. 4,794,175 and WO 88/09795); Talaromyces (U.S.Pat. No. 4,247,637; U.S. Pat. No. 6,255,084 and U.S. Pat. No.6,620,924); Rhizopus (Ashikari et al., (1986) Agric. Biol. Chem.50:957-964; Ashikari et al., (1989) App. Microbiol. Biotech. 32:129-133and U.S. Pat. No. 4,863,864); Humicola (WO 05/052148 and U.S. Pat. No.4,618,579) and Mucor (Houghton-Larsen et al., (2003) Appl. Microbiol.Biotechnol. 62:210-217). Many of the genes that code for these enzymeshave been cloned and expressed in yeast, fungal and/or bacterial cells.

Commercially, glucoamylases are very important enzymes and have beenused in a wide variety of applications that require the hydrolysis ofstarch (e.g., for producing glucose and other monosaccharides fromstarch). Glucoamylases are used to produce high fructose cornsweeteners, which comprise over 50% of the sweetener market in theUnited States. In general, glucoamylases may be, and commonly are, usedwith alpha amylases in starch hydrolyzing processes to hydrolyze starchto dextrins and then glucose. The glucose may then be converted tofructose by other enzymes (e.g., glucose isomerases); crystallized; orused in fermentations to produce numerous end products (e.g., ethanol,citric acid, lactic acid, succinate, ascorbic acid intermediates,glutamic acid, glycerol and 1,3-propanediol). Ethanol produced by usingglucoamylases in the fermentation of starch and/or cellulose containingmaterial may be used as a source of fuel or for alcoholic consumption.

Although glucoamylases have been used successfully in commercialapplications for many years, a need still exists for new glucoamylaseswith altered properties, such as improved specific activity andincreased thermostability.

Different mutations have been made in glucoamylases of Aspergillus thatenhance thermal stability and specific activity. Reference is made toU.S. Pat. No. 6,537,792; U.S. Pat. No. 6,352,851; Chen et al. (1996)Prot. Eng. 9:499-505, Chen et al., (1995) Prot Eng. 8:575-582; Fierobeet al. (1996) Biochem. 35:8698-8704; and Li et al., (1997) Prot. Eng.10:1199-1204. The need still exists for providing glucoamylase variantswith altered properties relative to their parent.

SUMMARY

The present disclosure relates to glucoamylase variants of a parentglucoamylase. The glucoamylase variants contain amino acid substitutionswith in the catalytic domain and/or the starch-binding domain. Thevariants display altered properties, such as improved thermostabilityand/or specific activity.

In one aspect, the present disclosure relates to a glucoamylase variantcomprising two or more amino acid substitutions corresponding toposition 61, 73, 417, 430, 431, 503, 511, 535, 539, or 563 of SEQ ID NO:2, or an equivalent position in a parent glucoamylase. In anotheraspect, the present disclosure relates a glucoamylase variant having atleast 80%, 85%, 90%, 95%, or 99.5% sequence identity with a parentglucoamylase of SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9. In one embodiment,the parent glucoamylase has a catalytic domain with at least 80%sequence identity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9, or a starchbinding domain with at least 80% sequence identity with SEQ ID NO: 1 or2. In other aspect, the parent glucoamylase is SEQ ID NO: 1 or 2. Afurther aspect of the present disclosure relates to glucoamylase variantfurther comprising one or more amino acid substitutions corresponding toposition: 4, 5, 12, 24, 43, 44, 45, 46, 47, 49, 51, 70, 75, 6, 94, 100,108, 114, 116, 119, 122, 124, 125, 137, 141, 143, 146, 148, 169, 171,172, 175, 178, 180, 181, 208, 211, 228, 242, 243, 245, 292, 294, 197,309, 310, 313, 314, 315, 316, 317, 321, 340, 341, 350, 353, 356, 363,368, 369, 375, 376, 395, 398, 401, 408, 409, 412, 415, 418, 421, 433,436 or 451 of SEQ ID NO: 2, or an equivalent position in the parentglucoamylase. In some aspects, the glucoamylase variant furthercomprises one or more amino acid substitutions corresponding toposition: 4, 5, 24, 29, 43, 44, 49, 70, 75, 76, 100, 108, 119, 124, 137,146, 148, 169, 171, 172, 175, 178, 181, 208, 211, 243, 292, 294, 297,314, 316, 317, 340, 341, 350, 356, 363, 368, 369, 376, 395, 401, 412,433, 436 or 451 of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase. In some aspects, the glucoamylase further comprises one ormore amino acid substitutions corresponding to position: 5, 24, 43, 44,49, 70, 75, 76, 94, 119, 141, 146, 148, 172, 175, 178, 180, 181, 208,211, 243, 294, 309, 314, 353, 369, 375, or 409 of SEQ ID NO: 2, or anequivalent position in a parent glucoamylase. In some aspects, theglucoamylase further comprises one or more amino acid substitutionscorresponding to position: 43, 44, or 294 of SEQ ID NO: 2, or anequivalent position in a parent glucoamylase.

In further aspects of the invention, the glucoamylase variant comprisestwo or more amino acid substitutions corresponding to position: N61I,G73F, L417R/V, T430A/M, A431L/Q, E503A/V, Q511H, A535R, A539R, orN563I/K of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase. In some aspects, the glucoamylase variant furthercomprises one or more of the following substitutions: D4L/E/R/S/C/A/Q/W,F5C/M/N/R/S/T/V/W, I12L/R, D24E/L/Y/T, F29L/I/D/C/S/V/W, I43F/R/D/Y/S/Q,D44E/H/K/S/N/Y/F/R/C, Y47W, Y49N, Q70R/K/M/P/G/L/F, Q75R/K/A,R76L/M/K/T/P, P94L, D100W/I/Q/M/P/A/N, N119P/T/Y/D/E,N146S/G/C/H/E/D/T/W/L/F/M, Q148V/Y/H/A/C/D/G/M/R/S/T, Y169D/F,Q172C/A/D/R/E/F/H/V/L/M/N/S/T/V, F175H/A/G/R/S/T/C/W/Y,W178A/C/D/E/F/G/H/K/N/R/S/T/V/Y, E180A/C/G/H/I/L/N/P/Q/R/S/T/V/Y/,V181E/C/D/G/H/I/P/T/Y/S/L/K/F/A, Q208L/A/C/E1N/F/H/T, S211C/R/E/A/Y/W/M/H/L/I/R/Q/T, E243 S/R/N/M/Y/A/L, R245A/E/M/I/P/V,I292D/H/P/R/T/N/V/F/L, G294C/D/E/T/Q/I/A,K297F/L/P/T/M/D/N/Q/A/Y/H/S/R/W, R309A/C/G/H/I/N/P/Q/S/T/W/Y/L,Y310E/G/L/P/S/W/R/Q, D313Q, V314A/R/N/D/C/E/Q/G/H/I/L/K/M/F/P/S/T/W/Y,Y315F, Y316Q/R, N317T/H, K340D/T, K341F/D/P/V/G/S, T350S/E/A/N,Q356H/D/E, T363L/R/C/H/W, S368W/D/F/L, S369F, N376Q/T/U/S/V, Y395Q/R/S,A398S/I/T, S401C/V, R408S, N409W/T/K, T412A/H/K/G, R433H/Q, I436A/T, orS451M/T/H of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase. In some aspects, the glucoamylase variant furthercomprises one or more of the following substitutions: I43F/R/D/Y/S/Q,D44E1H/K/S/N/Y/F/R/C, or G294C/D/E/T/Q/I/A of SEQ ID NO: 2, or anequivalent position in a parent glucoamylase.

In one aspect, the present disclosure relates a variant glucoamylasecomprises comprising amino acid substitutions corresponding topositions: I43Q/D44C, D44C/G294C, I43Q/G294C, or I43Q/D44C/G294 of SEQID NO: 2, or an equivalent position in a parent glucoamylase. Theglucoamylase variant has at least 80%, 85%, 90%, 95%, or 99.5% sequenceidentity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9. In one embodiment,the parent glucoamylase has a catalytic domain with at least 80%sequence identity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9, or a starchbinding domain with at least 80% sequence identity with SEQ ID NO: 1 or2.

The parent glucoamylase can the enzyme obtained from any of: aTrichoderma spp., an Aspergillus spp., a Humicola spp., a Penicilliumspp., a Talaromyces spp., or a Schizosaccharomyces spp. In some aspects,the parent glucoamylase can be from a Trichoderma spp. or an Aspergillusspp.

In one aspect, the variant glucoamylase exhibits altered thermostabilityas compared to the parent glucoamylase. The altered thermostability canbe increased thermostability. Alternatively, or in addition, the variantexhibits altered specific activity compared to the parent glucoamylase.The altered specific activity can be increased specific activity.

A further aspect of the disclosure is a polynucleotide encoding thevariant described. A further aspect is a vector comprising thepolynucleotide. A further aspect is a host cell containing the vector.

A further aspect of the disclosure is an enzyme composition includingthe glucoamylase variant. In one aspect, the enzyme composition is usedin a starch conversion process or an alcohol fermentation process.

A further aspect of the invention is a method of producing a variantglucoamylase by culturing the host cell containing the polynucleotideunder conditions suitable for the expression and production of theglucoamylase variant and producing the variant. The method may alsoinclude the step of recovering the glucoamylase variant from theculture.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1A depicts the Trichoderma reesei glucoamylase (TrGA) parentglucoamylase having 632 amino acids (SEQ ID NO: 1). The signal peptideis underlined, the catalytic region (SEQ ID NO: 3) starting with aminoacid residues SVDDFI (SEQ ID NO: 160) and having 453 amino acid residuesis in bold; the linker region is in italics; and the starch bindingdomain (SEQ ID NO: 161) is both italics and underlined. The matureprotein, which includes the catalytic domain (SEQ ID NO: 3), linkerregion, and starch binding domain (SEQ ID NO: 161), is represented bySEQ ID NO: 2. FIG. 1B depicts the cDNA (SEQ ID NO: 4) that codes for theTrGA.

FIG. 2 depicts the plasmid pDONR-TrGA that includes the cDNA (SEQ ID NO:4) of the parent TrGA.

FIG. 3 depicts the plasmids pREP3Y-DEST (A) and pREP3Y-TrGA (B).

FIGS. 4A-4B depict an alignment comparison of the catalytic domains ofparent glucoamylases including glucoamylase derived from Aspergillusawamori (AaGA) (SEQ ID NO: 5); Aspergillus niger (AnGA) (SEQ ID NO: 6);Aspergillus orzyae (AoGA) (SEQ ID NO: 7); Trichoderma reesei (TrGA) (SEQID NO: 3); Humicola grisea (HgGA) (SEQ ID NO: 8); and Hypocrea vinosa(HvGA) (SEQ ID NO: 9). Identical amino acids are indicated by anasterisk (*).

FIGS. 4C-4D depict an alignment comparing the Starch Binding Domain(SBD) of parent glucoamylases including Trichoderma reesei (TrGA) (SEQID NO: 161), Humicola grisea (HgGA) (SEQ ID NO: 162), Thielaviaterrestris (TtGA) (SEQ ID NO: 163), Thermomyces lanuginosus (ThGA) (SEQID NO: 164), Talaromyces emersonii (TeGA) (SEQ ID NO: 165), Aspergillusniger (AnGA) (SEQ ID NO: 166), and Aspergillus awamori (AaGA) (SEQ IDNO: 167). Identical amino acids are indicated by a dot (.).

FIG. 5A depicts the plasmid pTrex3g-DEST. FIG. 5B depicts the plasmidpTrex3g-TrGA. The plasmids were used as expression vectors forexpression and production of variant glucoamylases in a Trichodermareesei host.

FIG. 6 depicts the V_(max) (μM glucose/sec) comparison between theparent (wild-type) TrGA and variants, V314, S211R, Q172F, and Q208N at60° C. and 32° C. as further discussed in Example 8.

FIG. 7 depicts the activity of combinatorial variants on a starchsubstrate. Combinatorial variants described herein include ET7-1(D24Y/V181 L/Q208C/G294A/T353R/N375N/N409W), LR8 (Q172F/Q208N), LR12(Q172F/S211R), LR6 (Q172F/Q208N/V314H), ET8-1 (D24E/V181K/E243Y/I292V/G294Q/N409K), and ET7-2(Q24L/V181L/Q208C/G294A/T353R/N375Q/N409W). Activity is represented inunits of absorption at 340 nm as a function of ng of indicatedglucoamylase variants.

FIG. 8 depicts the activity of single site variants on a cornstarchsubstrate. Single site variants described herein include V314H, G294Q,S211R, Q208N, Q172F, G294I, and P94N. Activity is represented in unitsof absorption at 340 nm as a function of ng of indicated glucoamylasevariants.

FIG. 9 depicts the glucoamylase activity of TrGA and the TrGA variantLR8 (Q172F/Q208N) on a sample of corn mash liquefact (NE).

FIG. 10 depicts the activity profile of TrGA and the TrGA variant LR8(Q172F/Q208N) on a sample of corn mash liquefact (BSE).

FIG. 11 depicts the activity profile of TrGA and the TrGA variant LR8(Q172F/Q208N) on a sample soluble cornstarch substrate.

FIG. 12 depicts a comparison of the three dimensional structures ofTrichoderma reesei glucoamylase (black) (SEQ ID NO: 2) and Aspergillusawamorii glucoamylase (grey) viewed from the side.

FIG. 13 depicts a comparison of the three dimensional structures ofTrichoderma reesei glucoamylase (black) and Aspergillus awamoriiglucoamylase (grey) viewed from the top.

FIG. 14A depicts the plasmid pTTT-Dest. FIG. 14B depicts the plasmidpTTT-TrGA(B).

DETAILED DESCRIPTION

Glucoamylases are commercially important enzymes in a wide variety ofapplications that require the hydrolysis of starch. Glucoamylasesvariants of described herein contains amino acid substitutions withinthe catalytic domain or the starch binding domain. The variants maydisplay altered properties such as improved thermostability and/orspecific activity. The variants with improved thermostability and/orspecific activity may significantly improve the efficiency of glucoseand fuel ethanol production from corn starch, for example.

1. DEFINITIONS

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this disclosure belongs. Singleton, et al., DICTIONARYOF MICROBIOLOGY AND MOLECULAR BIOLOGY, 2D ED., John Wiley and Sons, NewYork (1994), and Hale & Markham, THE HARPER COLLINS DICTIONARY OFBIOLOGY, Harper Perennial, N.Y. (1991) provide one of skill with thegeneral meaning of many of the terms used herein. Certain terms aredefined below for the sake of clarity and ease of reference.

As used herein, the term “glucoamylase (EC 3.2.1.3)” refers to an enzymethat catalyzes the release of D-glucose from the non-reducing ends ofstarch and related oligo- and polysaccharides.

The term “parent” or “parent sequence” refers to a sequence that isnative or naturally occurring in a host cell. Parent sequences include,but are not limited to, the glucoamylase sequences set forth in SEQ IDNOs: 1, 2, 3, 5, 6, 7, 8, and 9.

As used herein, an “equivalent position” means a position that is commonto two parent sequences that is based on an alignment of the amino acidsequence of the parent glucoamylase in question as well as alignment ofthe three-dimensional structure of the parent glucoamylase in questionwith the TrGA reference glucoamylase amino acid sequence (SEQ ID NO: 2)and three-dimensional sequence.

The term “TrGA” refers to a parent Trichoderma reesei glucoamylasesequence having the mature protein sequence illustrated in SEQ ID NO: 2that includes the catalytic domain having the sequence illustrated inSEQ ID NO: 3. The isolation, cloning and expression of the TrGA aredescribed in U.S. Pat. No. 7,413,887, which are incorporated herein byreference. In some embodiments, the parent sequence refers to aglucoamylase that is the starting point for protein engineering. Thenumbering of the glucoamylase amino acids herein is based on thesequence alignment of a glucoamylase with TrGA (SEQ ID NO: 2 and SEQ IDNO: 3).

The phrase “mature form of a protein or polypeptide” refers to the finalfunctional form of the protein or polypeptide. To exemplify, a matureform of the TrGA includes the catalytic domain, linker region, andstarch binding domain having the amino acid sequence of SEQ ID NO: 2.

As used herein, the terms “glucoamylase variant” and “variant” are usedin reference to glucoamylases that have some degree of amino acidsequence identity to a parent glucoamylase sequence and that may retainthe functional characteristics of a glucoamylase. A variant is similarto a parent sequence, but has at least one substitution, deletion orinsertion in their amino acid sequence that makes them different insequence from a parent glucoamylase. In some cases, variants have beenmanipulated and/or engineered to include at least one substitution,deletion, or insertion in their amino acid sequence that makes themdifferent in sequence from a parent.

“Variants” may have at least 99.5%, at least 99%, at least 98%, at least97%, at least 96%, at least 95%, at least 94%, at least 93%, at least92%, at least 91%, at least 90%, at least 88%, at least 85%, at least80%, at least 75%, at least 70%, at least 65%, at least 60%, at least55%, at least 50%, or at least 45% sequence identity to a polypeptidesequence when optimally aligned for comparison. In some embodiments, theglucoamylase variant may have at least 99.5%, at least 99%, at least98%, at least 97%, at least 96%, at least 95%, at least 94%, at least93%, at least 92%, at least 91%, at least 90%, at least 88%, at least85%, at least 80%, at least 75%, at least 70%, at least 65%, at least60%, at least 55%, at least 50%, or at least 45% sequence identity tothe catalytic domain of a parent glucoamylase. In some embodiments, theglucoamylase variant may have at least 99.5%, at least 99%, at least98%, at least 97%, at least 96%, at least 95%, at least 94%, at least93%, at least 92%, at least 91%, at least 90%, at least 88%, at least85%, at least 80%, at least 75%, at least 70%, at least 65%, at least60%, at least 55%, at least 50%, or at least 45% sequence identity tothe starch binding domain of a parent glucoamylase. The sequenceidentity can be measured over the entire length of the parent or thevariant sequence.

Sequence identity is determined using standard techniques known in theart (see e.g., Smith and Waterman, Adv. Appl. Math. 2: 482 (1981);Needleman and Wunsch, J. Mol. Biol. 48: 443 (1970); Pearson and Lipman,Proc. Natl. Acad. Sci. USA 85: 2444 (1988); programs such as GAP,BESTHT, FASTA, and TFASTA in the Wisconsin Genetics Software Package(Genetics Computer Group, Madison, Wis.); and Devereux et al., NucleicAcid Res., 12: 387-395 (1984)).

The “percent (%) nucleic acid sequence identity” or “percent (%) aminoacid sequence identity” is defined as the percentage of nucleotideresidues or amino acid residues in a candidate sequence that areidentical with the nucleotide residues or amino acid residues of thestarting sequence (e.g., TrGA). The sequence identity can be measuredover the entire length of the starting sequence (e.g., SEQ ID NO: 2)

Sequence identity is determined by known methods of sequence alignment.A commonly used alignment method is BLAST described by Altschul et al.,(Altschul et al., J. Mol. Biol. 215: 403-410 (1990); and Karlin et al,Proc. Natl. Acad. Sci. USA 90: 5873-5787 (1993)). A particularly usefulBLAST program is the WU-BLAST-2 program (see Altschul et al, Meth.Enzymol. 266: 460-480 (1996)). WU-BLAST-2 uses several searchparameters, most of which are set to the default values. The adjustableparameters are set with the following values: overlap span=1, overlapfraction=0.125, word threshold (T)=11. The HSP S and HSP S2 parametersare dynamic values and are established by the program itself dependingupon the composition of the particular sequence and composition of theparticular database against which the sequence of interest is beingsearched. However, the values may be adjusted to increase sensitivity. A% amino acid sequence identity value is determined by the number ofmatching identical residues divided by the total number of residues ofthe “longer” sequence in the aligned region. The “longer” sequence isthe one having the most actual residues in the aligned region (gapsintroduced by WU-Blast-2 to maximize the alignment score are ignored).

Other methods find use in aligning sequences. One example of a usefulalgorithm is PILEUP. PILEUP creates a multiple sequence alignment from agroup of related sequences using progressive, pair-wise alignments. Itcan also plot a tree showing the clustering relationships used to createthe alignment. PILEUP uses a simplification of the progressive alignmentmethod of Feng and Doolittle (Feng and Doolittle, J. Mol. Evol. 35:351-360 (1987)). The method is similar to that described by Higgins andSharp (Higgins and Sharp, CABIOS 5: 151-153 (1989)). Useful PILEUPparameters including a default gap weight of 3.00, a default gap lengthweight of 0.10, and weighted end gaps.

The term “optimal alignment” refers to the alignment giving the highestpercent identity score.

As used herein the term “catalytic domain” refers to a structural regionof a polypeptide, which contains the active site for substratehydrolysis.

The term “linker” refers to a short amino acid sequence generally havingbetween 3 and 40 amino acids residues that covalently bind an amino acidsequence comprising a starch binding domain with an amino acid sequencecomprising a catalytic domain.

The term “starch binding domain” refers to an amino acid sequence thatbinds preferentially to a starch substrate.

As used herein, the terms “mutant sequence” and “mutant gene” are usedinterchangeably and refer to a polynucleotide sequence that has analteration in at least one codon occurring in a host cell's parentsequence. The expression product of the mutant sequence is a variantprotein with an altered amino acid sequence relative to the parent. Theexpression product may have an altered functional capacity (e.g.,enhanced enzymatic activity).

The term “property” or grammatical equivalents thereof in the context ofa polypeptide, as used herein, refers to any characteristic or attributeof a polypeptide that can be selected or detected. These propertiesinclude, but are not limited to oxidative stability, substratespecificity, catalytic activity, thermal stability, pH activity profile,resistance to proteolytic degradation, K_(M), K_(CAT), K_(CAT)/K_(M)ratio, protein folding, ability to bind a substrate and ability to besecreted.

The term “property” of grammatical equivalent thereof in the context ofa nucleic acid, as used herein, refers to any characteristic orattribute of a nucleic acid that can be selected or detected. Theseproperties include, but are not limited to, a property affecting genetranscription (e.g., promoter strength or promoter recognition), aproperty affecting RNA processing (e.g., RNA splicing and RNAstability), a property affecting translation (e.g., regulation, bindingof mRNA to ribosomal proteins).

The terms “thermally stable” and “thermostable” refer to glucoamylasevariants of the present disclosure that retain a specified amount ofenzymatic activity after exposure to a temperature over a given periodof time under conditions prevailing during the hydrolysis of starchsubstrates, for example, while exposed to altered temperatures.

The term “enhanced stability” in the context of a property such asthermostability refers to a higher retained starch hydrolytic activityover time as compared to another reference (i.e., parent) glucoamylases.

The term “diminished stability” in the context of a property such asthermostability refers to a lower retained starch hydrolytic activityover time as compared to another reference glucoamylase.

The term “specific activity” is defined as the activity per mg ofglucoamylase protein. In some embodiments, the activity for glucoamylaseis determined by the ethanol assay described herein and expressed as theamount of glucose that is produced from the starch substrate. In someembodiments, the protein concentration can be determined using theCaliper assay described herein.

The terms “active” and “biologically active” refer to a biologicalactivity associated with a particular protein. It follows that thebiological activity of a given protein refers to any biological activitytypically attributed to that protein by those skilled in the art. Forexample, an enzymatic activity associated with a glucoamylase ishydrolytic and, thus an active glucoamylase has hydrolytic activity.

The terms “polynucleotide” and “nucleic acid”, used interchangeablyherein, refer to a polymeric form of nucleotides of any length, eitherribonucleotides or deoxyribonucleotides. These terms include, but arenot limited to, a single-, double- or triple-stranded DNA, genomic DNA,cDNA, RNA, DNA-RNA hybrid, or a polymer comprising purine and pyrimidinebases, or other natural, chemically, biochemically modified, non-naturalor derivatized nucleotide bases.

As used herein, the terms “DNA construct,” “transforming DNA” and“expression vector” are used interchangeably to refer to DNA used tointroduce sequences into a host cell or organism. The DNA may begenerated in vitro by PCR or any other suitable technique(s) known tothose in the art. The DNA construct, transforming DNA or recombinantexpression cassette can be incorporated into a plasmid, chromosome,mitochondrial DNA, plastid DNA, virus, or nucleic acid fragment.Typically, the recombinant expression cassette portion of an expressionvector, DNA construct or transforming DNA includes, among othersequences, a nucleic acid sequence to be transcribed and a promoter. Insome embodiments, expression vectors have the ability to incorporate andexpress heterologous DNA fragments in a host cell.

As used herein, the term “vector” refers to a polynucleotide constructdesigned to introduce nucleic acids into one or more cell types. Vectorsinclude cloning vectors, expression vectors, shuttle vectors, plasmids,cassettes, and the like.

As used herein in the context of introducing a nucleic acid sequenceinto a cell, the term “introduced” refers to any method suitable fortransferring the nucleic acid sequence into the cell. Such methods forintroduction include but are not limited to protoplast fusion,transfection, transformation, conjugation, and transduction.

As used herein, the terms “transformed” and “stably transformed” refersto a cell that has a non-native (heterologous) polynucleotide sequenceintegrated into its genome or as an episomal plasmid that is maintainedfor at least two generations.

As used herein, the terms “selectable marker” and “selective marker”refer to a nucleic acid (e.g., a gene) capable of expression in hostcells that allows for ease of selection of those hosts containing thevector. Typically, selectable markers are genes that conferantimicrobial resistance or a metabolic advantage on the host cell toallow cells containing the exogenous DNA to be distinguished from cellsthat have not received any exogenous sequence during the transformation.

As used herein, the term “promoter” refers to a nucleic acid sequencethat functions to direct transcription of a downstream gene. Thepromoter, together with other transcriptional and translationalregulatory nucleic acid sequences (also termed “control sequences”) isnecessary to express a given gene. In general, the transcriptional andtranslational regulatory sequences include, but are not limited to,promoter sequences, ribosomal binding sites, transcriptional start andstop sequences, translational start and stop sequences, and enhancer oractivator sequences.

A nucleic acid is “operably linked” when it is placed into a functionalrelationship with another nucleic acid sequence. For example, DNAencoding a secretory leader (i.e., a signal peptide), is operably linkedto DNA for a polypeptide if it is expressed as a preprotein thatparticipates in the secretion of the polypeptide. Generally, “operablylinked” means that the DNA sequences being linked are contiguous, and,in the case of a secretory leader, contiguous and in reading phase.

As used herein the term “gene” refers to a polynucleotide (e.g., a DNAsegment), that encodes a polypeptide and includes regions preceding andfollowing the coding regions, as well as intervening sequences (introns)between individual coding segments (exons).

As used herein, “ortholog” and “orthologous genes” refer to genes indifferent species that have evolved from a common ancestral gene (i.e.,a homologous gene) by speciation. Typically, orthologs retain the samefunction during the course of evolution. Identification of orthologsfinds use in the reliable prediction of gene function in newly sequencedgenomes.

As used herein, “paralog” and “paralogous genes” refer to genes that arerelated by duplication within a genome. While orthologs retain the samefunction through the course of evolution, paralogs evolve new functions,even though some functions are often related to the original one.Examples of paralogous genes include, but are not limited to genesencoding trypsin, chymotrypsin, elastase, and thrombin, which are allserine proteinases and occur together within the same species.

As used herein, the term “hybridization” refers to the process by whicha strand of nucleic acid joins with a complementary strand through basepairing, as known in the art.

A nucleic acid sequence is considered to be “selectively hybridizable”to a reference nucleic acid sequence if the two sequences specificallyhybridize to one another under moderate to high stringency hybridizationand wash conditions. Hybridization conditions are based on the meltingtemperature (T_(m)) of the nucleic acid binding complex or probe. Forexample, “maximum stringency” typically occurs at about T_(m)−5° C. (5°below the T_(m) of the sequences”) is necessary to express a given gene.In general, the transcriptional and translational regulatory sequencesinclude, but are not limited to, promoter sequences, ribosomal bindingsites, transcriptional start and stop sequences, translational start andstop sequences, and enhancer or activator sequences.

A nucleic acid is “operably linked” when it is placed into a functionalrelationship with another nucleic acid sequence. For example, DNAencoding a secretory leader (i.e., a signal peptide), is operably linkedto DNA for a polypeptide if it is expressed as a preprotein thatparticipates in the secretion of the polypeptide. Generally, “operablylinked” means that the DNA sequences being linked are contiguous, and,in the case of a secretory leader, contiguous and in reading phase.

As used herein the term “gene” refers to a polynucleotide (e.g., a DNAsegment), that encodes a polypeptide and includes regions preceding andfollowing the coding regions, as well as intervening sequences (introns)between individual coding segments (exons).

As used herein, “ortholog” and “orthologous genes” refer to genes indifferent species that have evolved from a common ancestral gene (i.e.,a homologous gene) by speciation. Typically, orthologs retain the samefunction during the course of evolution. Identification of orthologsfinds use in the reliable prediction of gene function in newly sequencedgenomes.

As used herein, “paralog” and “paralogous genes” refer to genes that arerelated by duplication within a genome. While orthologs retain the samefunction through the course of evolution, paralogs evolve new functions,even though some functions are often related to the original one.Examples of paralogous genes include, but are not limited to genesencoding trypsin, chymotrypsin, elastase, and thrombin, which are allserine proteinases and occur together within the same species.

As used herein, the term “hybridization” refers to the process by whicha strand of nucleic acid joins with a complementary strand through basepairing, as known in the art.

A nucleic acid sequence is considered to be “selectively hybridizable”to a reference nucleic acid sequence if the two sequences specificallyhybridize to one another under moderate to high stringency hybridizationand wash conditions. Hybridization conditions are based on the meltingtemperature (T_(m)) of the nucleic acid binding complex or probe. Forexample, “maximum stringency” typically occurs at about T_(m)−5° C. (5°below the T_(m) of the probe); “high stringency” at about 5-10° C. belowthe T_(m); “intermediate stringency” at about 10-20° C. below the T_(m)of the probe; and “low stringency” at about 20-25° C. below the T_(m).Functionally, maximum stringency conditions may be used to identifysequences having strict identity or near-strict identity with thehybridization probe; while an intermediate or low stringencyhybridization can be used to identify or detect polynucleotide sequencehomologs.

Moderate and high stringency hybridization conditions are well known inthe art. An example of high stringency conditions includes hybridizationat about 42° C. in 50% formamide, 5×SSC, 5×Denhardt's solution, 0.5% SDSand 100 μg/ml denatured carrier DNA followed by washing two times in2×SSC and 0.5% SDS at room temperature and two additional times in0.1×SSC and 0.5% SDS at 42° C. An example of moderate stringentconditions include an overnight incubation at 37° C. in a solutioncomprising 20% formamide, 5×SSC (150 mM NaCl, 15 mM trisodium citrate),50 mM sodium phosphate (pH 7.6), 5×Denhardt's solution, 10% dextransulfate and 20 mg/ml denatured sheared salmon sperm DNA, followed bywashing the filters in 1×SSC at about 37-50° C. Those of skill in theart know how to adjust the temperature, ionic strength, etc. asnecessary to accommodate factors such as probe length and the like.

As used herein, “recombinant” includes reference to a cell or vector,that has been modified by the introduction of a heterologous orhomologous nucleic acid sequence or that the cell is derived from a cellso modified. Thus, for example, recombinant cells express genes that arenot found in identical form within the native (non-recombinant) form ofthe cell or express native genes that are otherwise abnormallyexpressed, under expressed or not expressed at all as a result ofdeliberate human intervention.

In an embodiment of the disclosure, mutated DNA sequences are generatedwith site saturation mutagenesis in at least one codon. In anotherembodiment, site saturation mutagenesis is performed for two or morecodons. In a further embodiment, mutant DNA sequences have more than50%, more than 55%, more than 60%, more than 65%, more than 70%, morethan 75%, more than 80%, more than 85%, more than 90%, more than 95%, ormore than 98% homology with the parent sequence. In alternativeembodiments, mutant DNA is generated in vivo using any known mutagenicprocedure such as, for example, radiation, nitrosoguanidine, and thelike. The desired DNA sequence is then isolated and used in the methodsprovided herein.

As used herein, “heterologous protein” refers to a protein orpolypeptide that does not naturally occur in the host cell.

An enzyme is “over-expressed” in a host cell if the enzyme is expressedin the cell at a higher level than the level at which it is expressed ina corresponding wild-type cell.

The terms “protein” and “polypeptide” are used interchangeabilityherein. In the present disclosure and claims, the conventionalone-letter and three-letter codes for amino acid residues are used. The3-letter code for amino acids as defined in conformity with theIUPAC-IUB Joint Commission on Biochemical Nomenclature (JCBN). It isalso understood that a polypeptide may be coded for by more than onenucleotide sequence due to the degeneracy of the genetic code.

Variants of the disclosure are described by the following nomenclature:[original amino acid residue/position/substituted amino acid residue].For example, the substitution of leucine for arginine at position 76 isrepresented as R76L. When more than one amino acid is substituted at agiven position, the substitution is represented as 1) Q 172C, Q172D orQ172R; 2) Q172C, D, or R, or 3) Q172C/D/R. When a position suitable forsubstitution is identified herein without a specific amino acidsuggested, it is to be understood that any amino acid residue may besubstituted for the amino acid residue present in the position. Where avariant glucoamylase contains a deletion in comparison with otherglucoamylases the deletion is indicated with “*”. For example, adeletion at position R76 is represented as R76*. A deletion of two ormore consecutive amino acids is indicated for example as (76-78)*.

A “prosequence” is an amino acid sequence between the signal sequenceand mature protein that is necessary for the secretion of the protein.Cleavage of the pro sequence will result in a mature active protein.

The term “signal sequence” or “signal peptide” refers to any sequence ofnucleotides and/or amino acids that may participate in the secretion ofthe mature or precursor forms of the protein. This definition of signalsequence is a functional one, meant to include all those amino acidsequences encoded by the N-terminal portion of the protein gene, whichparticipate in the effectuation of the secretion of protein. They areoften, but not universally, bound to the N-terminal portion of a proteinor to the N-terminal portion of a precursor protein. The signal sequencemay be endogenous or exogenous. The signal sequence may be that normallyassociated with the protein (e.g., glucoamylase), or may be from a geneencoding another secreted protein.

The term “precursor” form of a protein or peptide refers to a matureform of the protein having a prosequence operably linked to the amino orcarbonyl terminus of the protein. The precursor may also have a “signal”sequence operably linked, to the amino terminus of the prosequence. Theprecursor may also have additional polynucleotides that are involved inpost-translational activity (e.g., polynucleotides cleaved therefrom toleave the mature form of a protein or peptide).

“Host strain” or “host cell” refers to a suitable host for an expressionvector comprising DNA according to the present disclosure.

The terms “derived from” and “obtained from” refer to not only aglucoamylase produced or producible by a strain of the organism inquestion, but also a glucoamylase encoded by a DNA sequence isolatedfrom such strain and produced in a host organism containing such DNAsequence. Additionally, the term refers to a glucoamylase that isencoded by a DNA sequence of synthetic and/or cDNA origin and that hasthe identifying characteristics of the glucoamylase in question.

A “derivative” within the scope of this definition generally retains thecharacteristic hydrolyzing activity observed in the wild-type, native orparent form to the extent that the derivative is useful for similarpurposes as the wild-type, native or parent form. Functional derivativesof glucoamylases encompass naturally occurring, synthetically orrecombinantly produced peptides or peptide fragments that have thegeneral characteristics of the glucoamylases of the present disclosure.

The term “isolated” refers to a material that is removed from thenatural environment if it is naturally occurring.

A “purified” protein refers to a protein that is at least partiallypurified to homogeneity. In some embodiments, a purified protein is morethan 10% pure, optionally more than 20% pure, and optionally more than30% pure, as determined by SDS-PAGE. Further aspects of the disclosureencompass the protein in a highly purified form (i.e., more than 40%pure, more than 60% pure, more than 80% pure, more than 90% pure, morethan 95% pure, more than 97% pure, and even more than 99% pure), asdetermined by SDS-PAGE.

As used herein, the term, “combinatorial mutagenesis” refers to methodsin which libraries of variants of a starting sequence are generated. Inthese libraries, the variants contain one or several mutations chosenfrom a predefined set of mutations. In addition, the methods providemeans to introduce random mutations that were not members of thepredefined set of mutations. In some embodiments, the methods includethose set forth in U.S. Pat. No. 6,582,914, hereby incorporated byreference. In alternative embodiments, combinatorial mutagenesis methodsencompass commercially available kits (e.g., QuikChange® Multisite,Stratagene, San Diego, Calif.).

As used herein, the term “library of mutants” refers to a population ofcells that are identical in most of their genome but include differenthomologues of one or more genes. Such libraries can be used, forexample, to identify genes or operons with improved traits.

As used herein the term “dry solids content (DS or ds)” refers to thetotal solids of a slurry in % on a dry weight basis.

As used herein, the term “initial hit” refers to a variant that wasidentified by screening a combinatorial consensus mutagenesis library.In some embodiments, initial hits have improved performancecharacteristics, as compared to the starting gene.

As used herein, the term “improved hit” refers to a variant that wasidentified by screening an enhanced combinatorial consensus mutagenesislibrary.

As used herein, the term “target property” refers to the property of thestarting gene that is to be altered. It is not intended that the presentdisclosure be limited to any particular target property. However, insome embodiments, the target property is the stability of a gene product(e.g., resistance to denaturation, proteolysis or other degradativefactors), while in other embodiments, the level of production in aproduction host is altered. Indeed, it is contemplated that any propertyof a starting gene will find use in the present disclosure. Otherdefinitions of terms may appear throughout the specification.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimits of that range is also specifically disclosed. Each smaller rangebetween any stated value or intervening value in a stated range and anyother stated or intervening value in that stated range is encompassedwithin the disclosure. The upper and lower limits of these smallerranges may independently be included or excluded in the range, and eachrange where either, neither or both limits are included in the smallerranges is also encompassed within the disclosure, subject to anyspecifically excluded limit in the stated range. Where the stated rangeincludes one or both of the limits, ranges excluding either or both ofthose included limits are also included in the disclosure.

Before the exemplary embodiments are described in more detail, it is tobe understood that this disclosure is not limited to particularembodiments described, as such may, of course, vary. Although anymethods and materials similar or equivalent to those described hereincan be used in the practice or testing of the present disclosure,exemplary methods and materials are now described.

As used herein and in the appended claims, the singular forms “a”, “an”,and “the” include plural referents unless the context clearly dictatesotherwise. Thus, for example, reference to “a gene” includes a pluralityof such candidate agents and reference to “the cell” includes referenceto one or more cells and equivalents thereof known to those skilled inthe art, and so forth.

The publications discussed herein are provided solely for theirdisclosure prior to the filing date of the present application. Nothingherein is to be construed as an admission that the present disclosure isnot entitled to antedate such publication by virtue of prior invention.

2. PARENT GLUCOAMYLASES

In some embodiments, the present disclosure provides a glucoamylasevariant. The glucoamylase variant is a variant of a parent glucoamylase,which may comprise both a catalytic domain and a starch binding domain.In some embodiments, the parent glucoamylase comprises a catalyticdomain having an amino acid sequence as illustrated in SEQ ID NO: 1, 2,3, 5, 6, 7, 8 or 9 or having an amino acid sequence displaying at least80% sequence identity with one or more of the amino acid sequencesillustrated in SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9. In yet otherembodiments, the parent glucoamylase comprises a catalytic domainencoded by a DNA sequence that hybridizes under medium, high, orstringent conditions with a DNA encoding the catalytic domain of aglucoamylase having one of the amino acid sequences of SEQ ID NO: 1, 2or 3.

In some embodiments, the parent glucoamylase comprises a starch bindingdomain having an amino acid sequence as illustrated in SEQ ID NO 1, 2,161, 162, 163, 164, 165, 166, or 167, or having an amino acid sequencedisplaying at least 80% sequence identity with one or more of the aminoacid sequence illustrated in SEQ ID NO: 1, 2, 161, 162, 163, 164, 165,166, or 167. In yet other embodiments, the parent glucoamylase comprisesa starch binding domain encoded by a DNA sequence that hybridizes undermedium, high, or stringent conditions with a DNA encoding the starchbinding domain of a glucoamylase having one of the amino acid sequencesof SEQ ID NO: 1 or 2.

Predicted structure and known sequences of glucoamylases are conservedamong fungal species (Coutinho et al., 1994, Protein Eng., 7:393-400 andCoutinho et al., 1994, Protein Eng., 7: 749-760). In some embodiments,the parent glucoamylase is a filamentous fungal glucoamylase. In someembodiments, the parent glucoamylase is obtained from a Trichodermastrain (e.g., T. reesei, T. longibrachiatum, T. strictipilis, T.asperellum, T. konilangbra and T. hazianum), an Aspergillus strain (e.g.A. niger, A. nidulans, A. kawachi, A. awamori and A. orzyae), aTalaromyces strain (e.g. T. emersonii, T. thermophilus, and T. duponti),a Hypocrea strain (e.g. H. gelatinosa, H. orientalis, H. vinosa, and H.citrina), a Fusarium strain (e.g., F. oxysporum, F. roseum, and F.venenatum), a Neurospora strain (e.g., N. crassa) and a Humicola strain(e.g., H. grisea, H. insolens and H. lanuginose), a Penicillium strain(e.g., P. notatum or P. chrysogenum), or a Saccharomycopsis strain(e.g., S. fibuligera).

In some embodiments, the parent glucoamylase may be a bacterialglucoamylase. For example, the polypeptide may be obtained from agram-positive bacterial strain such as Bacillus (e.g., B. alkalophilus,B. amyloliquefaciens, B. lentus, B. licheniformis, B.stearothermophilus, B. subtilis and B. thuringiensis) or a Streptomycesstrain (e.g., S. lividans).

In some embodiments, the parent glucoamylase will comprise a catalyticdomain having at least 80% sequence identity, at least 85% sequenceidentity, at least 90% sequence identity, at least 95% sequenceidentity, at least 97% sequence identity and also at least 98% sequenceidentity with the catalytic domain of the TrGA amino acid sequence ofSEQ ID NO: 3.

In other embodiments, the parent glucoamylase will comprise a catalyticdomain having at least 90% sequence identity, at least 93% sequenceidentity, at least 95% sequence identity, at least 96% sequenceidentity, at least 97% sequence identity, at least 98% sequence identityand also at least 99% sequence identity with the catalytic domain of theAspergillus parent glucoamylase of SEQ ID NO: 5 or SEQ ID NO: 6.

In yet other embodiments, the parent glucoamylase will comprise acatalytic domain having at least 90% sequence identity, at least 95%sequence identity, at least 97% sequence identity and also at least 99%sequence identity with the catalytic domain of the Humicola grisea(HgGA) parent glucoamylase of SEQ ID NO: 8.

In some embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 80% sequence identity, at least 85%sequence identity, at least 90% sequence identity, at least 95% sequenceidentity, at least 97% sequence identity and also at least 98% sequenceidentity with the starch binding domain of the TrGA amino acid sequenceof SEQ ID NO: 1, 2, or 161.

In other embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 90% sequence identity, at least 95%sequence identity, at least 97% sequence identity and also at least 99%sequence identity with the catalytic domain of the Humicola grisea(HgGA) glucoamylase of SEQ ID NO: 162.

In other embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 90% sequence identity, at least 95%sequence identity, at least 97% sequence identity and also at least 99%sequence identity with the catalytic domain of the Thielavia terrestris(TtGA) glucoamylase of SEQ ID NO: 163.

In other embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 90% sequence identity, at least 95%sequence identity, at least 97% sequence identity and also at least 99%sequence identity with the catalytic domain of the Thermomyceslanuginosus (ThGA) glucoamylase of SEQ ID NO: 164.

In other embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 90% sequence identity, at least 95%sequence identity, at least 97% sequence identity and also at least 99%sequence identity with the catalytic domain of the Talaromycesemersoniit (TeGA) glucoamylase of SEQ ID NO: 165.

In yet other embodiments, the parent glucoamylase will comprise a starchbinding domain having at least 90% sequence identity, at least 93%sequence identity, at least 95% sequence identity, at least 96% sequenceidentity, at least 97% sequence identity, at least 98% sequence identityand also at least 99% sequence identity with the starch binding domainof the Aspergillus parent glucoamylase of SEQ ID NO: 166 or 167.

In some embodiments, the parent glucoamylase will have at least 80%sequence identity, at least 85% sequence identity, at least 88% sequenceidentity, at least 90% sequence identity, at least 93% sequenceidentity, at least 95% sequence identity, at least 96% sequenceidentity, at least 97% sequence identity, at least 98% sequence identityand also at least 99% sequence identity with the TrGA amino acidsequence of SEQ ID NO: 2.

In further embodiments, a Trichoderma glucoamylase homologue will beobtained from a Trichoderma or Hypocrea strain. Some typical Trichodermaglucoamylase homologues are described in U.S. Pat. No. 7,413,887 andreference is made specifically to amino acid sequences set forth in SEQID NOs: 17-22 and 43-47 of the reference.

In some embodiments, the parent glucoamylase is TrGA comprising theamino acid sequence of SEQ ID NO: 2, or a Trichoderma glucoamylasehomologue having at least 80%, at least 85%, at least 88%, at least 90%,at least 93%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% sequence identity to the TrGA sequence (SEQ ID NO: 2).

A parent glucoamylase can be isolated and/or identified using standardrecombinant DNA techniques. Any standard techniques can be used that areknown to the skilled artisan. For example, probes and/or primersspecific for conserved regions of the glucoamylase can be used toidentify homologues in bacterial or fungal cells (the catalytic domain,the active site, etc.). Alternatively, degenerate PCR can be used toidentify homologues in bacterial or fungal cells. In some cases, knownsequences, such as in a database, can be analyzed for sequence and/orstructural identity to one of the known glucoamylases, including SEQ IDNO: 2, or a known starch binding domains, including SEQ ID NO: 161.Functional assays can also be used to identify glucoamylase activity ina bacterial or fungal cell. Proteins having glucoamylase activity can beisolated and reverse sequenced to isolate the corresponding DNAsequence. Such methods are known to the skilled artisan.

3. GLUCOAMYLASE STRUCTURAL HOMOLOGY

The central dogma of molecular biology is that the sequence of DNAencoding a gene for a particular enzyme, determines the amino acidsequence of the protein, this sequence in turn determines thethree-dimensional folding of the enzyme. This folding brings togetherdisparate residues that create a catalytic center and substrate bindingsurface and this results in the high specificity and activity of theenzymes in question.

Glucoamylases consist of as many as three distinct structural domains, acatalytic domain of approximately 450 residues that is structurallyconserved in all glucoamylases, generally followed by a linker regionconsisting of between 30 and 80 residues that are connected to a starchbinding domain of approximately 100 residues. The structure of theTrichoderma reesei glucoamylase with all three regions intact wasdetermined to 1.8 Angstrom resolution herein (see Table 20 and Example13). Using the coordinates (see Table 20) the structure was aligned withthe coordinates of the catalytic domain from Aspergillus awamorii strainX100 that was determined previously (Aleshin, A. E., Hoffman, C.,Firsov, L. M., and Honzatko, R. B. 1994 Refined crystal structures ofglucoamylase from Aspergillus awamori var. X100. J Mol Biol 238:575-591.). The Aspergillus awamori crystal structure only included thecatalytic domain. As seen in FIGS. 12 and 13, the structure of thecatalytic domains overlap very closely and it is possible to identifyequivalent residues based on this structural superposition. It isbelieved that all glucoamylases share the basic structure depicted inFIGS. 12 and 13.

FIG. 12 is a comparison of the three dimensional structures of theTrichoderma glucoamylase (black) of SEQ ID NO: 1 (see FIG. 1 for aminoacid sequence) and of Aspergillus awamorii (grey) viewed from the side.In this view the relationship between the catalytic domain and thelinker region and the starch binding domain can be seen.

FIG. 13 is a comparison of the three dimensional structures of theTrichoderma glucoamylase (black) and of Aspergillus awamorii (grey)viewed from the top. The glucoamylases shown here and indeed all knownglucoamylases to date share this structural homology. The conservationof structure correlates with the conservation of activity and aconserved mechanism of action for all glucoamylases. Given this highhomology, changes resulting from site specific variants of theTrichoderma glucoamylase resulting in altered function would also havesimilar structural and therefore functional consequences in otherglucoamylases. Therefore, the teachings of which variants result indesirable benefits can be applied to other glucoamylases.

A further crystal structure was produced using the coordinates in Table20 for the Starch Binding Domain (SBD). The SBD for TrGA was alignedwith the SBD for A. niger. As shown in FIG. 13, the structure of the A.niger and TrGA SBDs overlaps very closely. It is believed that while allstarch binding domains share at least some of the basic structuredepicted in FIG. 13, some SBDs are more structurally similar thanothers. For example, the TrGA SBD can be classified as within thecarbohydrate binding module 20 family within the CAZY database(cazy.org). The CAZY database describes the families ofstructurally-related catalytic and carbohydrate-binding modules (orfunctional domains) of enzymes that degrade, modify, or createglycosidic bonds. Given a high structural homology, site specificvariants of the TrGA SBD resulting in altered function would also havesimilar structural and therefore functional consequences in otherglucoamylases having SBDs with similar structure to that of the TrGASBD, particularly those classified within the carbohydrate bindingmodule 20 family. Thus, the teachings of which variants result indesirable benefits can be applied to other SBDs having structuralsimilarity.

Thus, the amino acid position numbers discussed herein refer to thoseassigned to the mature Trichoderma reesei glucoamylase sequencepresented in FIG. 1. The present disclosure, however, is not limited tothe variants of Trichoderma glucoamylase, but extends to glucoamylasescontaining amino acid residues at positions that are “equivalent” to theparticular identified residues in Trichoderma reesei glucoamylase (SEQID NO: 2). In a embodiment of the present disclosure, the parentglucoamylase is Taleromyces glucoamylase and the substitutions are madeat the equivalent amino acid residue positions in Taleromycesglucoamylase as those described herein. In other embodiments, the parentglucoamylase is one of those listed in Table 1. In further embodiments,the parent glucoamylase is a Penicillium glucoamylase, such asPenicillium chrysogenum,

Structural identity determines whether the amino acid residues areequivalent. Structural identity is a one-to-one topological equivalentwhen the two structures (three dimensional and amino acid structures)are aligned. A residue (amino acid) position of a glucoamylase isequivalent to a residue of T. reesei glucoamylase if it is eitherhomologous (i.e., corresponding in position in either primary ortertiary structure) or analogous to a specific residue or portion ofthat residue in T. reesei glucoamylase (having the same or similarfunctional capacity to combine, react, or interact chemically).

In order to establish identity to the primary structure, the amino acidsequence of a glucoamylase can be directly compared to Trichodermareesei glucoamylase primary sequence and particularly to a set ofresidues known to be invariant in glucoamylases for which sequence isknown. For example, FIGS. 4A and 4B herein shows the conserved residuesbetween glucoamylases. FIGS. 4C and 4D show an alignment of starchbinding domains from various glucoamylases. After aligning the conservedresidues, allowing for necessary insertions and deletions in order tomaintain alignment (i.e. avoiding the elimination of conserved residuesthrough arbitrary deletion and insertion), the residues equivalent toparticular amino acids in the primary sequence of Trichoderma reeseiglucoamylase are defined. Alignment of conserved residues typicallyshould conserve 100% of such residues. However, alignment of greaterthan 75% or as little as 50% of conserved residues is also adequate todefine equivalent residues. Further, the structural identity can be usedin combination with the sequence identity to identify equivalentresidues.

For example, in FIGS. 4A and 4B, the catalytic domains of glucoamylasesfrom six organisms are aligned to provide the maximum amount of homologybetween amino acid sequences. A comparison of these sequences shows thatthere are a number of conserved residues contained in each sequence asdesignated by an asterisk. These conserved residues, thus, may be usedto define the corresponding equivalent amino acid residues ofTrichoderma reesei glucoamylase in other glucoamylases such asglucoamylase from Aspergillus niger. Similarly, FIGS. 4C and 4D show thestarch binding domains of glucoamylases from seven organisms aligned toidentify equivalent residues.

Structural identity involves the identification of equivalent residuesbetween the two structures. “Equivalent residues” can be defined bydetermining homology at the level of tertiary structure (structuralidentity) for an enzyme whose tertiary structure has been determined byX-ray crystallography. Equivalent residues are defined as those forwhich the atomic coordinates of two or more of the main chain atoms of aparticular amino acid residue of the Trichoderma reesei glucoamylase (Non N, CA on CA, C on C and O on O) are within 0.13 nm and optionally 0.1nm after alignment. Alignment is achieved after the best model has beenoriented and positioned to give the maximum overlap of atomiccoordinates of non-hydrogen protein atoms of the glucoamylase inquestion to the Trichoderma reesei glucoamylase. The best model is thecrystallographic model giving the lowest R factor for experimentaldiffraction data at the highest resolution available.

${R\mspace{14mu} {factor}} = \frac{{\Sigma_{h}{{{Fo}(h)}}} - {{{Fc}(h)}}}{\Sigma_{h}{{{Fo}(h)}}}$

Equivalent residues that are functionally analogous to a specificresidue of Trichoderma reesei glucoamylase are defined as those aminoacids of the enzyme that may adopt a conformation such that they eitheralter, modify or contribute to protein structure, substrate binding orcatalysis in a manner defined and attributed to a specific residue ofthe Trichoderma reesei glucoamylase. Further, they are those residues ofthe enzyme (for which a tertiary structure has been obtained by X-raycrystallography) that occupy an analogous position to the extent that,although the main chain atoms of the given residue may not satisfy thecriteria of equivalence on the basis of occupying a homologous position,the atomic coordinates of at least two of the side chain atoms of theresidue lie with 0.13 nm of the corresponding side chain atoms ofTrichoderma reesei glucoamylase. The coordinates of the threedimensional structure of Trichoderma reesei glucoamylase are set forthin Table 15 and can be used as outlined above to determine equivalentresidues on the level of tertiary structure.

Some of the residues identified for substitution are conserved residueswhereas others are not. In the case of residues that are not conserved,the substitution of one or more amino acids is limited to substitutionsthat produce a variant that has an amino acid sequence that does notcorrespond to one found in nature. In the case of conserved residues,such substitutions should not result in a naturally-occurring sequence.

4. VARIANTS

The variants according to the disclosure include at least onesubstitution, deletion or insertion in the amino acid sequence of aparent glucoamylase that makes the variant different in sequence fromthe parent glucoamylase. In some embodiments, the variants of thedisclosure will have at least 20%, at least 40%, at least 60%, at least80%, at least 85%, at least 90%, at least 95%, or at least 100% of theglucoamylase activity of the TrGA activity of SEQ ID NO: 2.

In some embodiments, the variants according to the disclosure willcomprise a substitution, deletion, or insertion in at least one aminoacid position of the parent TrGA (SEQ ID NO: 2), or in an equivalentposition in the sequence of another parent glucoamylase having at least80% sequence identity to the TrGA sequence, including but not limitedto, at least 90%, at least 93%, at least 95%, at least 97%, and at least99% sequence identity.

In other embodiments, the variant according to the disclosure willcomprise a substitution, deletion or insertion in at least one aminoacid position of a fragment of the parent TrGA, wherein the fragmentcomprises the catalytic domain of the TrGA sequence (SEQ ID NO: 3) or inan equivalent position in a fragment comprising the catalytic domain ofa parent glucoamylase having at least 80% sequence identity to thefragment of the TrGA sequence, including but not limited to, at least90%, at least 95%, at least 97%, and at least 99%. In some embodiments,the fragment will comprise at least 400, 425, 450, and/or 500 amino acidresidues.

In other embodiments, the variant according to the disclosure willcomprise a substitution, deletion or insertion in at least one aminoacid position of a fragment of the parent TrGA, wherein the fragmentcomprises the starch binding domain of the TrGA sequence (SEQ ID NO:161) or in an equivalent position in a fragment comprising the starchbinding domain of a parent glucoamylase having at least 80% sequenceidentity to the fragment of the TrGA sequence, including but not limitedto, at least 90%, at least 95%, at least 97%, and at least 99%. In someembodiments, the fragment will comprise at least 40, 50, 60, 70, 80, 90,100, and/or 109 amino acid residues of TrGA starch binding domain (SEQID NO: 161).

In some embodiments, when the parent glucoamylase includes a catalyticdomain, a linker region, and a starch binding domain, the variant willcomprise a substitution, deletion or insertion in at least one aminoacid position of a fragment comprising part of the linker region. Insome embodiments, the variant will comprise a substitution deletion, orinsertion in the amino acid sequence of a fragment of the TrGA sequence(SEQ ID NO: 2).

Structural identity with reference to an amino acid substitution meansthat the substitution occurs at the equivalent amino acid position inthe homologous glucoamylase or parent glucoamylase. The term equivalentposition means a position that is common to two parent sequences that isbased on an alignment of the amino acid sequence of the parentglucoamylase in question as well as alignment of the three-dimensionalstructure of the parent glucoamylase in question with the TrGA referenceglucoamylase amino acid sequence and three-dimensional sequence. Forexample, with reference to FIG. 5, position 24 in TrGA (SEQ ID NO: 2 or3) is D24 and the equivalent position for Aspergillus niger (SEQ ID NO:6) is position D25, and the equivalent position for Aspergillus oryzea(SEQ ID NO: 7) is position D26. See FIGS. 12 and 13 for an exemplaryalignment of the three-dimensional sequence.

In some embodiments, the glucoamylase variant will include at least onesubstitution in the amino acid sequence of a parent. In furtherembodiments, the variant may have more than one substitution (e.g., two,three, or four substitutions).

In some embodiments, a glucoamylase variant comprises a substitution,deletion or insertion, and typically a substitution in at least oneamino acid position in a position corresponding to the regions ofnon-conserved amino acids as illustrated in FIG. 5 (e.g., amino acidpositions corresponding to those positions that are not designated by“*” in FIG. 5).

While the variants can be in any position in the mature protein sequence(SEQ ID NO: 2), in one embodiment, a glucoamylase variant comprises oneor more substitutions in the following positions in the amino acidsequence set forth in SEQ ID NO: 2: 4, 5, 12, 24, 29, 43, 44, 45, 46,47, 49, 51, 61, 70, 73, 75, 76, 94, 100, 108, 114, 116, 119, 122, 124,125, 137, 143, 146, 148, 169, 171, 172, 175, 178, 180, 181, 208, 211,228, 242, 243, 245, 292, 294, 297, 309, 310, 313, 314, 315, 316, 317,321, 340, 341, 350, 353, 356, 363, 368, 369, 375, 376, 395, 398, 401,408, 409, 412, 415, 417, 418, 421, 430, 431, 433, 436, 451, 503, 511,535, 539, or 563; or in an equivalent position in a parent glucoamylase.In some embodiments, the parent glucoamylase will have at least 90%, atleast 95%, at least 96%, at least 97% at least 98%, and at least 99%identity with SEQ ID NO: 2. In other embodiments the parent glucoamylasewill be a Trichoderma glucoamylase homologue.

In some embodiments, the glucoamylase variant comprises one or moresubstitutions in the following positions in the amino acid sequence setforth in SEQ ID NO: 2:

D4, F5, I12, D24, F29, I43, D44, P45, D46, Y47, Y49, W51, N61, Y70, G73,Q75, R76, P94, D100, K108, K114, F116, N119, R122, Q124, R125, G137,N146, Q148, Y169, N171, Q172, F175, W178, E180, V181, Q208, S211, W228,N242, E243, R245, I292, G294, K297, R309, Y310, D313, V314, Y315, Y316,N317, W321, K340, K341, T350, Q356, T363, S368, S369, N376, Y395, A398,S401, R408, N409, T412, L417, H418, W421, T430, A431, R433, I436, S451,E503, Q511, A535, A539, or N563; or an equivalent position in parentglucoamylase (e.g., a Trichoderma glucoamylase homologue).

In other embodiments, the variant of a glucoamylase parent comprises oneor more substitutions in the following positions in the amino acidsequence set forth in SEQ ID NO:2: 4, 5, 24, 29, 43, 44, 49, 61, 70, 73,75, 76, 100, 108, 119, 124, 137, 146, 148, 169, 171, 172, 175, 178, 181,208, 211, 243, 292, 294, 297, 314, 316, 317, 340, 341, 350, 356, 363,368, 369, 376, 395, 401, 409, 412, 417, 430, 431, 433, 436, 451, 503,511, 535, 539, or 563; or an equivalent position in a parentglucoamylase (e.g., a Trichoderma glucoamylase homologue).

In further embodiments, the variant of a glucoamylase parent comprisesat least one of the following substitutions in the following positionsin an amino acid sequence set forth in SEQ ID NO: 2: D4L/E/R/S/C/A/Q/W,F5C/M/N/R/S/T/V/W, I12L/R, D24E/L/Y/T, F29L/I/D/C/S/V/W, I43F/R/D/Y/S/Q,D44E/H/K/S/N/Y/F/R/C, Y47W, Y49N, N61D/I/L/Q/V/W, Q70R/K/M/P/G/L/F,G73F/C/L/W, Q75R/K/A, R76L/M/K/T/P, P94L, D100W/I/Q/M/P/A/N,N119P/T/Y/D/E, N146S/G/C/H/E/D/T/W/L/F/M, Q148V/Y/H/A/C/D/G/M/R/S/T,Y169D/F, Q172C/A/D/R/E/F/H/V/L/M/N/S/T/V, F175H/A/G/R/S/T/C/W/Y,W178A/C/D/E/F/G/H/K/N/R/S/T/V/Y, E180A/C/G/H/I/L/N/P/Q/R/S/T/V/Y/,V181E/C/D/G/H/I/P/T/Y/S/L/K/F/A, Q208L/A/C/E/N/F/H/T,S211C/R/E/A/Y/W/M/H/L/I/R/Q/T, E243 S/R/N/M/Y/A/L, R245A/E/M/I/P/V,I292D/H/P/R/T/N/V/F/L, G294C/D/E/T/Q/I/A,K297F/L/P/T/M/D/N/Q/A/Y/H/S/R/W, R309A/C/G/H/I/N/P/Q/S/T/W/Y/L,Y310E/G/L/P/S/W/R/Q, D313Q, V314A/R/N/D/C/E/Q/G/H/I/L/K/M/F/P/S/T/W/Y,Y315F, Y316Q/R, N317T/H, K340D/T, K341F/D/P/V/G/S, T350S/E/A/N,Q356H/D/E, T363L/R/C/H/W, S368W/D/F/L, S369F, N376Q/T/H/S/V, Y395Q/R/S,A398S/I/T, S401C/V, R408S, N409W/T/K, T412A/H/K/G,L417A/D/E/F/G/I/K/Q/R/S/T/V/W/Y, T430A/E/F/G/H/I/K/M/N/Q/R/V,A431C/E/H/I/L/M/Q/R/S/W/Y, R433H/Q, I436A/T, S451M/T/H,E503A/C/D/H/S/V/W, Q511C/G/H/I/K/T/V, A535E/F/G/K/L/N/P/R/S/T/V/W/Y,A539E/H/M/R/S/W, or N563/A/C/E/I/K/L/Q/T/V; or a substitution in anequivalent position in a parent glucoamylase homologue.

In some embodiments, the glucoamylase variant comprises at least onesubstitution in a position corresponding to the amino acid residueposition set forth in SEQ ID NO: 2: 5, 24, 43, 44, 49, 61, 70, 73, 75,76, 94, 119, 146, 148, 172, 175, 178, 180, 181, 208, 211, 245, 294, 353,315, 375, 409, 309, 314, 369, 412, 417, 430, 431, 503, 511, 535, 539, or563; or an equivalent position in a homologous parent glucoamylase.

In some representative embodiments, the glucoamylase variant comprisesat least one substitution selected from the group consisting of F5W,D24E, I43R, I43Y, I43Q, I43S, I43F, D44C, D44R, Y47W, Y49N, N61I, Q70K,G73F, Q75R, R76L, P94L, N119P/T/Y/D, N146S/D/T/E/W/L, Q148V N171D,Q172C/D/R/E/F/V/L/T, F175R/W/Y, W178K/N/Y, E180H/N/V/R, V181E/F/G/I/H,Q208A/T/N, S211H/M/L/R, R245E, R245M, G294C, R309W, V314F/G/H/K/P/R/Y,Y315F, S369F, T412K, L417R, L417V, T430A, T430M, A431L, A431Q, E503A,E503V, Q511H, A535R, A539R, N563I, and N563K corresponding to theposition set forth in SEQ ID NO: 2, or an equivalent position in ahomologous parent glucoamylase.

In further particular embodiments, the glucoamylase variant comprises atleast one substitution of an amino acid residue selected from thepositions corresponding to position 5, 43, 44, 61, 73, 75, 76, 94, 108,119, 124, 146, 148, 171, 172, 175, 178, 180, 181, 208, 211, 294, 297,314, 316, 412, 417, 430, 431, 503, 511, 535, 539, or 563 of SEQ ID NO:2, or an equivalent position in a Trichoderma glucoamylase homologue. Insome embodiments, the substitution is at a position corresponding toposition number 43, 44, 61, 73, 148, 172, 175, 178, 180, 208, 211, 294,297, 314, 412, 417, 430, 431, 503, 511, 535, 539, or 563 of SEQ ID NO:2, or an equivalent position in a Trichoderma glucoamylase homologue.

In some representative embodiments, the substitution is at a positioncorresponding to position number 43, 44, 61, 73, 108, 124, 171, 172,208, 211, 294, 314, 316, 417, 430, 431, 503, 511, 535, 539, or 563 ofSEQ ID NO: 2, or a homologous parent glucoamylase (e.g., Trichodermaglucoamylase homologue).

In some embodiments, the glucoamylase variants comprise multiplesubstitutions. Some of the multiple substitutions will include asubstitution at one or more of the positions equivalent to and includingthe positions 24, 43, 44, 108, 124, 171, 175, 181, 208, 243, 292, 294,297, 310, 314, 363, 417, 430, 431, 503, 511, 535, 539, or 563 of SEQ IDNO: 2. Some typical multiple substitutions will include one or more ofthe positions equivalent to and corresponding to positions 43, 44, 61,73, 08, 124, 171, 208, 211, 294, 314 417, 430, 431, 503, 511, 535, 539,or 563 of SEQ ID NO: 2.

Some examples of variants with multiple substitutions includesubstitutions at positions:

D24/I43/D44/F175V181/V314/T363;

D24/Q208/I292/G294/K297/Y310;

V181/E243/I292/K297/N317/Y395;

D24/V181/Q208/G294/T363/N376/N409;

D24/V181/I292/G294/E243/N409;

I43R/E243/I292/G294/K297;

I43/D44/N61/L417/E503/Q511/A539;

I43/D44/L417/E503/Q511/A539;

I43/N61/L417/T430/Q511/A539;

I43/N61/L417/E503/Q511/A539;

I43/N61/T430/A431/Q511/A539;

I43/N61/T430/Q511;

I43/N61/T430/Q511/A539;

I43/N61/Q511;

I43/N61/Q511/A539;

I43/G73/T430;

I43/L417/E503/Q511/A539;

I43/L417/Q511;

I43/L417/T430/A431/Q511/A539;

I43/L417/T430/Q511;

I43/L417/T430/Q511/A539;

I43/L417/E503/A539;

I43/L417/E503/Q511/A539;

I43/T430;

I43/T430/A431/E503/Q511;

I43/T430/A431/Q511;

I43/T430/A431/Q511/A539;

I43/T430/E503/Q511;

I43/T430/Q511;

I43Q/T430/Q511/A539;

I43/A431/Q511;

I43/T430/E503/Q511/N563;

I43/T430/E503/A535/N563;

I43/E503/Q511/A539;

I43/Q511/A539;

D44/G73/L417/N563;

D44/G73/E503/Q511;

D44/G73/N563;

D44/L417/N563;

D44/T430/Q511/A535;

D44/E503/Q511/N563;

G73/T430/E503/Q511;

G73/T430/Q511;

G294/L417/A431;

G294/L417/A431;

G294/L417/A431/Q511;

L417/T430/A431/Q511/A535/A539/N563;

L417/A431/Q511;

L417/T430/Q511/A535/N563;

L417/T430/Q511/A539/N563; and

E503/N563;

of SEQ ID NO: 2, or equivalent positions in parent glucoamylases andparticularly Trichoderma glucoamylase homologues.

Some variants with multiple substitutions may include the substitutionsat positions:

Y47F/W,Y315F/W;

D24E,L/I43F,R/D44H,N/F175H/V181K,L/V314D,H,K/T363R;

D24L,W,Y/Q208F/I292F,N,V/G294A,I,Q/K297A/Y310F,Q,R;

V181F,K,L/E243A,N,M,R,Y/I292F,L,N,V/K297A,D,H,M,N,Q/N317H/Y395Q,R;

D24E,L,Y/V181F,K,L/Q208C,F/G294A,I,Q/T363R/N376Q/N409K, W;

D24E,L,Y/V181F,K,L/I292F,L,N,V/G294A,I,Q/E243A,M,N,R,Y/N409K, W;

I43R/E243A,M,N,R,Y/I292F,L,N,V/G294A/K297A,D,H,M,N,Q,S,R,W,Y;

I43Q/D44C/N611/L417V/E503A/Q511H/A539R;

I43Q/D44C/L417V/E503A/Q511H/A539R;

I43Q/N61I/L417V/E503A/Q511H/A539R;

I43Q/N61I/L417V/T430M/Q51H/A539R;

I43R/N61I/L417R,V/E503A/Q511H/A539R;

I43Q/N61I/T430A/A431L/Q511H/A539R;

I43Q/N61I/T430A/Q511H;

I43Q/N61I/T430A/Q511H/A539R;

I43Q/N61I/T430M/Q511H/A539R

I43Q/N61I/Q511H;

I43Q/N61I/Q511H/A539R;

I43R/G73F/T430A;

I43Q/L417V/T430A/A431L/Q511H/A539R;

I43Q/L417V/T430A/Q511H;

I43Q/L417V/T430A/Q511H/A539R;

I43R/L417R/E503A/A539R;

I43R,Q/L417V/E503A/Q511H/A539R;

I43Q/L417V/Q511H;

I43R,Q/T430A;

I43Q/T430A/A431L/E503A/Q511H;

I43Q/T430A/A431 L/Q511H;

I43Q/T430A/A431 L/Q511H/A539R;

I43Q/T430A/E503A/Q511H;

I43Q/T430A/Q511H;

I43Q/T430A,M/Q511H/A539R;

I43R/T430A/E503A,V/Q511H/N563K;

I43Q/A431L/Q511H;

I43Q/E503A/Q511H/A539R;

I43Q/Q511H/A539R;

D44C/G73F/E503V/Q511H;

D44C/G73F/L417R/N563K;

D44C/G73F/N563K;

D44C/L417R/N563K;

D44R/E503A/Q511H/N563I;

D44R/T430A/Q511H/A535R;

G73F/T430A/E503V/Q511H;

G73F/T430A/Q511H;

G294C/L417R/A431L;

G294C/L417R/A431 L,Q/Q511H;

G294C/L417V/A431Q;

L417R,V/A431 L,Q/Q511H;

L417V/T430A/A431L,Q/Q511H/A535R/A539R/N563I;

L417V/T430A/Q511H/A535R/N563I;

L417V/T430A/Q511H/A539R/N563I; and

E503A/N563I

of SEQ ID NO: 2, or equivalent positions in parent glucoamylases andparticularly Trichoderma glucoamylase homologue.

A number of parent glucoamylases have been aligned with the amino acidsequence of TrGA. FIGS. 4A and 4B include the catalytic domain of thefollowing parent glucoamylases: Aspergillus awamori (AaGA) (SEQ ID NO:5); Aspergillus niger (AnGA) (SEQ ID NO: 6); Aspergillus orzyae (AoGA)(SEQ ID NO: 7), Humicola grisea (HgGA) (SEQ ID NO: 8) and Hypocreavinosa (HvGA) (SEQ ID NO: 9). The percent identity of the catalyticdomains is represented in Table I below. FIGS. 4C and 4D include thestarch binding domain of the following parent glucoamylases: Trichodermareesei (TrGA) (SEQ ID NO: 161), Humicola grisea (HgGA) (SEQ ID NO: 162),Thielavia terrestris (TtGA) (SEQ ID NO: 163), Thermomyces lanuginosus(ThGA) (SEQ ID NO: 164), Talaromyces emersonii (TeGA) (SEQ ID NO: 165),Aspergillus niger (AnGA) (SEQ ID NO: 166), and Aspergillus awamori(AaGA) (SEQ ID NO: 167). In some embodiments, for example, the variantglucoamylase will be derived from a parent glucoamylase that is anAspergillus glucoamylase and the variant will include at least onesubstitution in a position equivalent to a position set forth in SEQ IDNO: 2, and particularly in a position corresponding to D4, F5, I12, D24,F29, I43, D44, P45, D46, Y47, Y49, W51, N61, Y70, G73, Q75, R76, P94,D100, K108, K114, F116, N119, R122, Q124, R125, G137, N146, Q148, Y169,N171, Q172, F175, W178, E180, V181, Q208, S211, W228, N242, E243, R245,I292, G294, K297, R309, Y310, D313, V314, Y315, Y316, N317, W321, K340,K341, T350, Q356, T363, S368, S369, N376, Y395, A398, S401, R408, N409,T412, L417, H418, W421, T430, A431, R433, I436, S451, E503, Q511, A535,A539, or N563.

Endo-H removal of N-linked sugars in the Trichoderma reesei glucoamylasehad a stabilizing effect (when looking at T_(m)). Thus, variants havingan N171D substitution can have increased thermostability as compared tothe wild-type parent. In some embodiments, variants having one or moresubstitutions at sites having N-linked sugars are provided, includingN171D in Trichoderma reesei (SEQ ID NO: 2).

TABLE 1 AaGA AnGA AoGA HgGA HvGA TrGA AaGA 100 95 58 53 57 56 AnGA 10059 53 57 56 AoGA 100 55 56 56 HgGA 100 61 63 HvGA 100 91 TrGA 100

The present disclosure also provides glucoamylase variants having atleast one altered property (e.g., improved property) as compared to aparent glucoamylase and particularly to the TrGA. In some embodiments,at least one altered property (e.g., improved property) is selected fromthe group consisting of acid stability, thermal stability and specificactivity. Typically, the altered property is increased acid stability,increased thermal stability and/or increased specific activity. Theincreased thermal stability typically is at higher temperatures. In oneembodiment, the increased pH stability is at high pH. In a furtherembodiment, the increased pH stability is at low pH.

The glucoamylase variants of the disclosure may also provide higherrates of starch hydrolysis at low substrate concentrations as comparedto the parent glucoamylase. The variant may have a higher V_(max) orlower K_(m) than a parent glucoamylase when tested under the sameconditions. For example the variant glucoamylase may have a higherV_(max) at a temperature range of 25° C. to 70° C. (e.g., at 25° C. to35° C.; 30° C. to 35° C.; 40° C. to 50° C.; at 50° C. to 55° C. and at55° C. to 62° C.). The Michaelis-Menten constant, K_(m) and V_(max)values can be easily determined using standard known procedures.

5. THERMAL STABILITY THERMOSTABLE VARIANTS

In one aspect, the disclosure relates to a variant glucoamylase havingaltered thermal stability at altered temperatures as compared to aparent or wild type. Altered temperatures include increased or decreasedtemperatures. In some embodiments, the glucoamylase variant will haveimproved thermostability such as retaining at least 50%, 60%, 70%, 75%,80%, 85%, 90%, 92%, 95%, 96%, 97%, 98% or 99% enzymatic activity afterexposure to altered temperatures over a given time period, for example,at least 60 minutes, 120 minutes, 180 minutes, 240 minutes, 300 minutes,etc. In some embodiments, the variant has increased thermal stabilitycompared to the parent glucoamylase at selected temperatures in therange of 40 to 80° C., also in the range of 50 to 75° C. and in therange of 60 to 70° C., and typically at a pH range of 4.0 to 6.0. Insome embodiments, the thermostability is determined as described in theExamples.

In some embodiments, particularly interesting variants in connectionwith an improvement in thermostability include one or more deletions,substitutions or insertions and particularly substitutions in thefollowing positions in the amino acid sequence set forth in SEQ ID NO:2: D4, F5, I12, D24, F29, I43, D44, P45, D46, Y47, Y49, W51, N61,Y70,G73, Q75, R76, P94, D100, K108, K114, F116, N119, R122, Q124, R125,G137, N146, Q148, Y169, N171, Q172, F175, W178, E180, V181, Q208, S211,W228, N242, E243, R245, I292, G294, K297, R309, Y310, D313, V314, Y315,Y316, N317, W321, K340, K341, T350, Q356, T363, S368, S369, N376, Y395,A398, S401, R408, N409, T412, L417, H418, W421, T430, A431, R433, I436,S451, E503, Q511, A535, A539, or N563; or an equivalent position in aparent glucoamylase. In some embodiments, the parent glucoamylase willbe a Trichoderma glucoamylase homologue, and in typical embodiments, theparent glucoamylase will have at least 90%, at least 95%, or at least98% sequence identity to SEQ ID NO: 2.

6. CHIMERIC GLUCOAMYLASES

Glucoamylase variants of the disclosure may also include chimeric orhybrid glucoamylases with, for example a starch binding domain (SBD)from one glucoamylase and a catalytic domain and linker from another.For example, a hybrid glucoamylase can be made by swapping the SBD fromAnGA with the SBD from TrGA, making a hybrid with the AnGA SBD and theTrGA catalytic domain and linker. Alternatively, the SBD and linker fromAnGA can be swapped for the SBD and linker of TrGA.

7. SPECIFIC ACTIVITY

In another aspect, the disclosure relates to a variant glucoamylasehaving altered specific activity as compared to a parent or wild-typeglucoamylase.

In some embodiments, particularly interesting variants in connectionwith an improvement in specific activity include one or more deletions,substitutions or insertions and particularly substitutions in thefollowing positions in the amino acid sequence set forth in SEQ ID NO:2: D4, F5, I12, D24, F29, I43, D44, P45, D46, Y47, Y49, W51, N61, Y70,G73, Q75, R76, P94, D100, K108, K114, F116, N119, R122, Q124, R125,G137, N146, Q148, Y169, N171, Q172, F175, W178, E180, V181, Q208, S211,W228, N242, E243, R245, I292, G294, K297, R309, Y310, D313, V314, Y315,Y316, N317, W321, K340, K341, T350, Q356, T363, S368, S369, N376, Y395,A398, S401, R408, N409, T412, L417, T430, A431, H418, W421, R433, I436,S451, E503, Q511, A535, A539, or N563; or an equivalent position in aparent glucoamylase. In some embodiments, variants of the disclosurehaving improved specific activity include a substitution in thefollowing positions in the amino acid sequence set forth in SEQ ID NO:2: D4, D24, I43, D44, N61, Y70, G73, Q75, R76, D100, K108, N119, Q124,N146, Q148, N171, Q172, F175, V181, Q208, S211, E243, R245, I292, G294,K297, V314, Y316, N317, K340, K341, T350, Q356, T363, S368, N376, Y395,A398, S401, N409, T412, L417, T430, A431, I436, S451, E503, Q511, A535,A539, or N563; or an equivalent position in a parent glucoamylase. Insome embodiments, the parent glucoamylase will comprise a sequencehaving at least 90% or 95% sequence identity to the sequence of SEQ IDNO: 2.

8. POLYNUCLEOTIDES

The present disclosure also relates to isolated polynucleotides encodinga variant glucoamylase of the disclosure. The polynucleotides encoding avariant glucoamylase may be prepared by established techniques known inthe art. The polynucleotides may be prepared synthetically, such as byan automatic DNA synthesizer. The DNA sequence may be of mixed genomic(or cDNA) and synthetic origin prepared by ligating fragments together.The polynucleotides may also be prepared by polymerase chain reaction(PCR) using specific primers. In general, reference is made to MinshullJ., et al., (2004), Engineered protein function by selective amino aciddiversification, Methods 32(4):416-427. Also a number of companies nowsynthesize DNA such as Geneart AG, Regensburg, Germany.

The present disclosure also provides isolated polynucleotides comprisinga nucleotide sequence (i) having at least 70% identity to SEQ ID NO: 4,or (ii) being capable of hybridizing to a probe derived from thenucleotide sequence set forth in SEQ ID NO: 4, under conditions ofintermediate to high stringency, or (iii) being complementary to anucleotide sequence having at least 90% sequence identity to thesequence set forth in SEQ ID NO: 4. Probes useful according to thedisclosure may include at least 50, 100, 150, 200, 250, 300 or morecontiguous nucleotides of SEQ ID NO: 4.

The present disclosure further provides isolated polynucleotides thatencode variant glucoamylases that comprise an amino acid sequencecomprising at least 80% amino acid sequence identity to SEQ ID NO: 2. Insome embodiments, the variant glucoamylases have at least 80% amino acidsequence identity to SEQ ID NO: 2. In some embodiments, the variantglucoamylases have at least 90% amino acid sequence identity to SEQ IDNO: 2. In some embodiments, the variant glucoamylases have at least 93%amino acid sequence identity to SEQ ID NO: 2. In some embodiments, thevariant glucoamylases have at least 95% amino acid sequence identity toSEQ ID NO: 2. In some embodiments, the variant glucoamylases have atleast 97% amino acid sequence identity to SEQ ID NO: 2. In someembodiments, the variant glucoamylases have at least 98% amino acidsequence identity to SEQ ID NO: 2. In some embodiments, the variantglucoamylases have at least 99% amino acid sequence identity to SEQ IDNO: 2. The present disclosure also provides expression vectorscomprising any of the polynucleotides provided above.

The present disclosure also provides fragments (i.e., portions) of theDNA encoding the variant glucoamylases provided herein. These fragmentsfind use in obtaining partial length DNA fragments capable of being usedto isolate or identify polynucleotides encoding mature glucoamylaseenzymes described herein from filamentous fungal cells (e.g.,Trichoderma, Aspergillus, Fusarium, Penicillium, Schizosaccharomyces,and Humicola), or a segment thereof having glucoamylase activity. Insome embodiments, fragments of the DNA may comprise at least 50, 100,150, 200, 250 300 or more contiguous nucleotides. In some embodiments,portions of the DNA provided in SEQ ID NO: 4 may be used in obtainingparent glucoamylase and particularly Trichoderma glucoamylase homologuesfrom other species, such as filamentous fungi that encode aglucoamylase.

9. DNA CONSTRUCTS AND VECTORS

According to one embodiment of the disclosure, a DNA constructcomprising a polynucleotide as described above encoding a variantglucoamylase encompassed by the disclosure and operably linked to apromoter sequence is assembled to transfer into a host cell.

The DNA construct may be introduced into a host cell using a vector. Thevector may be any vector that when introduced into a host cell istypically integrated into the host cell genome and is replicated.Vectors include cloning vectors, expression vectors, shuttle vectors,plasmids, phage particles, cassettes and the like. In some embodiments,the vector is an expression vector that comprises regulatory sequencesoperably linked to the glucoamylase coding sequence.

Examples of suitable expression and/or integration vectors are providedin Sambrook et al., (1989) supra, and Ausubel (1987) supra, and van denHondel et al. (1991) in Bennett and Lasure (Eds.) MORE GENEMANIPULATIONS IN FUNGI, Academic Press pp. 396-428 and U.S. Pat. No.5,874,276. Fungal Genetics Stock Center Catalogue of Strains (FGSC,<www.fgsc.net>) also discloses useful vectors. Particularly usefulvectors include vectors obtained from, for example, Invitrogen andPromega.

Specific vectors suitable for use in fungal host cells include vectorssuch as pFB6, pBR322, pUC18, pUC100, pDONR™201, pDONR™221, pENTR™,pGEM®3Z and pGEM®4Z. A general purpose expression vector useful inAspergillus includes pRAX with a glaA promoter, and inHypocrea/Trichoderma includes pTrex3g with a cbh1 promoter.

Suitable plasmids for use in bacterial cells include pBR322 and pUC19permitting replication in E. coli and pE194 for example permittingreplication in Bacillus.

In some embodiments, the promoter shows transcriptional activity in abacterial or a fungal host cell and may be derived from genes encodingproteins either homologous or heterologous to the host cell. Thepromoter may be a mutant, a truncated and/or a hybrid promoter. Theabove-mentioned promoters are known in the art.

Examples of suitable promoters useful in fungal cells and particularlyfilamentous fungal cells such as Trichoderma or Aspergillus cellsinclude such exemplary promoters as the T. reesei promoters cbh1, cbh2,egl1, egl2, eg5, xln1 and xln2. Other examples of useful promotersinclude promoters from A. awamori and A. niger glucoamylase genes (glaA)(see, Nunberg et al., (1984) Mol. Cell. Biol. 4:2306-2315 and Boel etal., (1984) EMBO J. 3:1581-1585), A. oryzae TAKA amylase promoter, theTPI (triose phosphate isomerase) promoter from S. cerevisiae, thepromoter from Aspergillus nidulans acetamidase genes and Rhizomucormiehei lipase genes.

Examples of suitable promoters useful in bacterial cells include thoseobtained from the E. coli lac operon; Bacillus licheniformis alphaamylase gene (amyL), B. stearothermophilus amylase gene (amyM); Bacillussubtilis xylA and xylB genes, the beta-lactamase gene, and the tacpromoter.

In one embodiment, the promoter is one that is native to the host cell.For example, when T. reesei is the host, the promoter is a native T.reesei promoter. In another embodiment, the promoter is one that isheterologous to the fungal host cell. In some embodiments the promoterwill be the parent glucoamylase promoter such as the TrGA promoter.

In some embodiments, the DNA construct includes nucleic acids coding fora signal sequence that is an amino acid sequence linked to the aminoterminus of the polypeptide that directs the encoded polypeptide intothe cell's secretory pathway. The 5′ end of the coding sequence of thenucleic acid sequence may naturally include a signal peptide codingregion that is naturally linked in translation reading frame with thesegment of the glucoamylase coding sequence that encodes the secretedglucoamylase or the 5′ end of the coding sequence of the nucleic acidsequence may include a signal peptide that is foreign to the codingsequence. In some embodiments, the DNA construct includes a signalsequence that is naturally associated with a parent glucoamylase genefrom that a variant glucoamylase has been obtained. In some embodimentsthe signal sequence will be the sequence depicted in SEQ ID NO: 1 or asequence having at least 90%, at least 94% and at least 98% sequenceidentity thereto. Effective signal sequences may include the signalsequences obtained from glucoamylases of other filamentous fungalenzymes, such as from Trichoderma (T. reesei glucoamylase), Humicola (H.insolens cellulase or H. grisea glucoamylase), Aspergillus (A. nigerglucoamylase and A. oryzae TAKA amylase), and Rhizopus.

In additional embodiments, a DNA construct or vector comprising a signalsequence and a promoter sequence to be introduced into a host cell arederived from the same source. In some embodiments, the nativeglucoamylase signal sequence of a Trichoderma glucoamylase homologue,such as a signal sequence from a Hypocrea strain may be used.

In some embodiments, the expression vector also includes a terminationsequence. Any terminator sequence functional in the host cell may beused in the present disclosure. In one embodiment, the terminationsequence and the promoter sequence are derived from the same source. Inanother embodiment, the termination sequence is homologous to the hostcell. Useful terminator sequence include terminator sequences obtainedfrom the genes of Trichoderma reesei cbh1; A. niger or A. awamoriglucoamylase (Nunberg et al. (1984) supra, and Boel et al., (1984)supra), Aspergillus nidulans anthranilate synthase, Aspergillus oryzaeTAKA amylase, or A. nidulans trpC (Punt et al., (1987) Gene 56:117-124).

In some embodiments, an expression vector includes a selectable marker.Examples of typical selectable markers include ones that conferantimicrobial resistance (e.g., hygromycin and phleomycin). Nutritionalselective markers also find use in the present disclosure includingthose markers known in the art as amdS (acetamidase), argB (ornithinecarbamoyltransferase) and pyrG (orotidine-5′phosphate decarboxylase).Markers useful in vector systems for transformation of Trichoderma areknown in the art (see, e.g., Finkelstein, chapter 6 in BIOTECHNOLOGY OFFILAMENTOUS FUNGI, Finkelstein et al. Eds. Butterworth-Heinemann,Boston, Mass. (1992); Kinghorn et al. (1992) APPLIED MOLECULAR GENETICSOF FILAMENTOUS FUNGI, Blackie Academic and Professional, Chapman andHall, London; Berges and Barreau (1991) Curr. Genet. 19:359-365; and vanHartingsveldt et al., (1987) Mol. Gen. Genet. 206:71-75). In a typicalembodiment, the selective marker is the amdS gene, which encodes theenzyme acetamidase, allowing transformed cells to grow on acetamide as anitrogen source. The use of A. nidulans amdS gene as a selective markeris described in Kelley et al., (1985) EMBO J. 4:475-479 and Penttilä etal., (1987) Gene 61:155-164.

Methods used to ligate the DNA construct comprising a nucleic acidsequence encoding a variant glucoamylase, a promoter, a terminator andother sequences and to insert them into a suitable vector are well knownin the art. Linking is generally accomplished by ligation at convenientrestriction sites. If such sites do not exist, synthetic oligonucleotidelinkers are used in accordance with conventional practice. (See,Sambrook (1989) supra, and Bennett and Lasure, MORE GENE MANIPULATIONSIN FUNGI, Academic Press, San Diego (1991) pp 70-76.). Additionally,vectors can be constructed using known recombination techniques (e.g.,Invitrogen Life Technologies, Gateway Technology).

10. HOST CELLS

The present disclosure also relates to host cells comprising apolynucleotide encoding a variant glucoamylase of the disclosure, whichare used to produce the glucoamylases of the disclosure. In someembodiments, the host cells are selected from bacterial, fungal, plantand yeast cells. The term host cell includes both the cells, progeny ofthe cells and protoplasts created from the cells that are used toproduce a variant glucoamylase according to the disclosure.

In some embodiments, the host cells are fungal cells and typicallyfilamentous fungal host cells. The term “filamentous fungi” refers toall filamentous forms of the subdivision Eumycotina (See, Alexopoulos,C. J. (1962), INTRODUCTORY MYCOLOGY, Wiley, New York). These fungi arecharacterized by a vegetative mycelium with a cell wall composed ofchitin, cellulose, and other complex polysaccharides. The filamentousfungi of the present disclosure are morphologically, physiologically,and genetically distinct from yeasts. Vegetative growth by filamentousfungi is by hyphal elongation and carbon catabolism is obligatoryaerobic. In the present disclosure, the filamentous fungal parent cellmay be a cell of a species of, but not limited to, Trichoderma, (e.g.,Trichoderma reesei, the asexual morph of Hypocrea jecorina, previouslyclassified as T. longibrachiatum, Trichoderma viride, Trichodermakoningii, Trichoderma harzianum) (Sheir-Neirs et al., (1984) Appl.Microbiol. Biotechnol 20:46-53; ATCC No. 56765 and ATCC No. 26921);Penicillium sp., Humicola sp. (e.g., H. insolens, H. lanuginosa and H.grisea); Chrysosporium sp. (e.g., C. lucknowense), Gliocladium sp.,Aspergillus sp. (e.g., A. oryzae, A. niger, A. sojae, A. japonicus, A.nidulans, and A. awamori) (Ward et al., (1993) Appl. Microbiol.Biotechnol. 39:738-743 and Goedegebuur et al., (2002) Genet. 41:89-98),Fusarium sp., (e.g., F. roseum, F. graminum F. cerealis, F. oxysporumand F. venenatum), Neurospora sp., (N. crassa), Hypocrea sp., Mucor sp.,(M. miehei), Rhizopus sp. and Emericella sp. (see also, Innis et al.,(1985) Sci. 228:21-26). The term “Trichoderma” or “Trichoderma sp.” or“Trichoderma spp.” refer to any fungal genus previously or currentlyclassified as Trichoderma.

In some embodiments, the host cells will be gram-positive bacterialcells. Non-limiting examples include strains of Streptomyces, (e.g., S.lividans, S. coelicolor and S. griseus) and Bacillus. As used herein,“the genus Bacillus” includes all species within the genus “Bacillus,”as known to those of skill in the art, including but not limited to B.subtilis, B. licheniformis, B. lentus, B. brevis, B. stearothermophilus,B. alkalophilus, B. amyloliquefaciens, B. clausii, B. halodurans, B.megaterium, B. coagulans, B. circulans, B. lautus, and B. thuringiensis.It is recognized that the genus Bacillus continues to undergotaxonomical reorganization. Thus, it is intended that the genus includespecies that have been reclassified, including but not limited to suchorganisms as B. stearothermophilus, which is now named “Geobacillusstearothermophilus.”

In some embodiments the host cell is a gram-negative bacterial strain,such as E. coli or Pseudomonas sp. In other embodiments, the host cellsmay be yeast cells such as Saccharomyces sp., Schizosaccharomyces sp.,Pichia sp., or Candida sp.

In other embodiments, the host cell will be a genetically engineeredhost cell wherein native genes have been inactivated, for example bydeletion in bacterial or fungal cells. Where it is desired to obtain afungal host cell having one or more inactivated genes known methods maybe used (e.g. methods disclosed in U.S. Pat. No. 5,246,853, U.S. Pat.No. 5,475,101, and WO 92/06209). Gene inactivation may be accomplishedby complete or partial deletion, by insertional inactivation or by anyother means that renders a gene nonfunctional for its intended purpose(such that the gene is prevented from expression of a functionalprotein). In some embodiments, when the host cell is a Trichoderma celland particularly a T. reesei host cell, the cbh1, cbh2, egl1 and egl2genes will be inactivated and/or typically deleted. Typically,Trichoderma reesei host cells having quad-deleted proteins are set forthand described in U.S. Pat. No. 5,847,276 and WO 05/001036. In otherembodiments, the host cell is a protease deficient or protease minusstrain.

11. TRANSFORMATION OF HOST CELLS

Introduction of a DNA construct or vector into a host cell includestechniques such as transformation; electroporation; nuclearmicroinjection; transduction; transfection, (e.g., lipofection mediatedand DEAE-Dextrin mediated transfection); incubation with calciumphosphate DNA precipitate; high velocity bombardment with DNA-coatedmicroprojectiles; and protoplast fusion. General transformationtechniques are known in the art (See, e.g., Ausubel et al., (1987),supra, chapter 9; and Sambrook (1989) supra, and Campbell et al., (1989)Curr. Genet. 16:53-56).

Transformation methods for Bacillus are disclosed in numerous referencesincluding Anagnostopoulos C and J. Spizizen (1961) J. Bacteriol.81:741-746 and WO 02/14490.

Transformation methods for Aspergillus are described in Yelton et al(1984) Proc. Natl. Acad. Sci. USA 81:1470-1474; Berka et al., (1991) inApplications of Enzyme Biotechnology, Eds. Kelly and Baldwin, PlenumPress (NY); Cao et al., (2000) Science 9:991-1001; Campbell et al.,(1989) Curr. Genet. 16:53-56 and EP 238 023. The expression ofheterologous protein in Trichoderma is described in U.S. Pat. No.6,022,725; U.S. Pat. No. 6,268,328; Harkki et al. (1991); Enzyme Microb.Technol. 13:227-233; Harkki et al., (1989) Bio Technol. 7:596-603; EP244,234; EP 215,594; and Nevalainen et al., “The Molecular Biology ofTrichoderma and its Application to the Expression of Both Homologous andHeterologous Genes”, in MOLECULAR INDUSTRIAL MYCOLOGY, Eds. Leong andBerka, Marcel Dekker Inc., NY (1992) pp. 129-148). Reference is alsomade to WO96/00787 and Bajar et al., (1991) Proc. Natl. Acad. Sci. USA88:8202-8212 for transformation of Fusarium strains.

In one specific embodiment, the preparation of Trichoderma sp. fortransformation involves the preparation of protoplasts from fungalmycelia (See, Campbell et al., (1989) Curr. Genet. 16:53-56; Pentilla etal., (1987) Gene 61:155-164). Agrobacterium tumefaciens-mediatedtransformation of filamentous fungi is known (See, de Groot et al.,(1998) Nat. Biotechnol. 16:839-842). Reference is also made to U.S. Pat.No. 6,022,725 and U.S. Pat. No. 6,268,328 for transformation proceduresused with filamentous fungal hosts.

Typically, genetically stable transformants are constructed with vectorsystems whereby the nucleic acid encoding the variant glucoamylase isstably integrated into a host strain chromosome. Transformants are thenpurified by known techniques.

In some further embodiments, the host cells are plant cells, such ascells from a monocot plant (e.g. corn, wheat and sorghum) or cells froma dicot plant (e.g. soybean). Methods for making DNA constructs usefulin transformation of plants and methods for plant transformation areknown. Some of these methods include Agrobacterium tumefaciens mediategene transfer; microprojectile bombardment, PEG mediated transformationof protoplasts, electroporation and the like. Reference is made to (U.S.Pat. No. 6,803,499, U.S. Pat. No. 6,777,589; Fromm et al (1990)Biotechnol. 8:833-839; Potrykus et al (1985) Mol. Gen. Genet.199:169-177.

12. PRODUCTION OF PROTEINS

The present disclosure further relates to methods of producing thevariant glucoamylases comprising transforming a host cell with anexpression vector comprising a polynucleotide encoding a variantglucoamylase according to the disclosure, optionally culturing the hostcell under conditions suitable for production of the variantglucoamylase and optionally recovering the glucoamylase.

In the expression and production methods of the present disclosure thehost cells are cultured under suitable conditions in shake flaskcultivation, small scale or large scale fermentations (includingcontinuous, batch and fed batch fermentations) in laboratory orindustrial fermentors, with suitable medium containing physiologicalsalts and nutrients (see, e.g., Pourquie, J. et al., BIOCHEMISTRY ANDGENETICS OF CELLULOSE DEGRADATION, eds. Aubert, J. P. et al., AcademicPress, pp. 71-86, 1988 and Ilmen, M. et al., (1997) Appl. Environ.Microbiol. 63:1298-1306). Common commercially prepared media (e.g.,Yeast Malt Extract (YM) broth, Luria Bertani (LB) broth and SabouraudDextrose (SD) broth) find use in the present disclosure. Representativeculture conditions for bacterial and filamentous fungal cells are knownin the art and may be found in the scientific literature and/or from thesource of the fungi such as the American Type Culture Collection andFungal Genetics Stock Center. In cases where a glucoamylase codingsequence is under the control of an inducible promoter, the inducingagent (e.g., a sugar, metal salt or antimicrobial), is added to themedium at a concentration effective to induce glucoamylase expression.

In some embodiments, the present disclosure relates to methods ofproducing the variant glucoamylase comprising cultivating a plant orplant cell comprising a polynucleotide encoding a variant glucoamylaseaccording to the disclosure under conditions suitable for the productionof the variant and optionally recovering the glucoamylase.

In some embodiments, in order to evaluate the expression of a variantglucoamylase by a cell line that has been transformed with apolynucleotide encoding a variant glucoamylase encompassed by thedisclosure, assays are carried out at the protein level, the RNA leveland/or by use of functional bioassays particular to glucoamylaseactivity and/or production. Some of these assays include Northernblotting, dot blotting (DNA or RNA analysis), RT-PCR (reversetranscriptase polymerase chain reaction), or in situ hybridization,using an appropriately labeled probe (based on the nucleic acid codingsequence) and conventional Southern blotting and autoradiography.

In addition, the production and/or expression of a variant glucoamylasemay be measured in a sample directly, for example, by assays directlymeasuring reducing sugars such as glucose in the culture medium and byassays for measuring glucoamylase activity, expression and/orproduction. In particular, glucoamylase activity may be assayed by the3,5-dinitrosalicylic acid (DNS) method (see, Goto et al., (1994) Biosci.Biotechnol. Biochem. 58:49-54). In additional embodiments, proteinexpression, is evaluated by immunological methods, such asimmunohistochemical staining of cells, tissue sections or immunoassay oftissue culture medium, (e.g., by Western blot or ELISA). Suchimmunoassays can be used to qualitatively and quantitatively evaluateexpression of a glucoamylase. The details of such methods are known tothose of skill in the art and many reagents for practicing such methodsare commercially available.

The glucoamylases of the present disclosure may be recovered or purifiedfrom culture media by a variety of procedures known in the art includingcentrifugation, filtration, extraction, precipitation and the like.

13. COMPOSITIONS

The variant glucoamylases may be used in enzyme compositions includingbut not limited to starch hydrolyzing and saccharifying compositions,cleaning and detergent compositions (e.g., laundry detergents, dishwashing detergents, and hard surface cleaning compositions), alcoholfermentation compositions, and in animal feed compositions. Further thevariant glucoamylases may be used in baking applications, such as breadand cake production, brewing, healthcare, textile, environmental wasteconversion processes, biopulp processing, and biomass conversionapplications.

In some embodiments, an enzyme composition including a variantglucoamylase encompassed by the disclosure obtained in culture media orrecovered and purified from the culture medium will be optionally usedin combination with any one or combination of the followingenzymes—alpha amylases, proteases, pullulanases, isoamylases,cellulases, hemicellulases, xylanases, cyclodextrin glycotransferases,lipases, phytases, laccases, oxidases, esterases, cutinases, xylanases,granular starch hydrolyzing enzyme and other glucoamylases. In oneembodiment, the proteases are acid fungal proteases (AFP). In a furtherembodiment, the acid fungal proteases are from Trichoderma (e.g.,NSP-24, see also US 2006/0154353, published Jul. 13, 2006, incorporatedherein by reference). In a further embodiment, the phytase is fromButtiauxiella. spp. (e.g., BP-17, see also variants disclosed in PCTpatent publication WO 2006/043178).

In some representative compositions, the variant glucoamylases of thedisclosure will be combined with an alpha amylase, such as fungal alphaamylases (e.g., Aspergillus sp.) or bacterial alpha amylases (e.g.,Bacillus sp. such as B. stearothermophilus, B. amyloliquefaciens and B.licheniformis) and variants and hybrids thereof. In one embodiment thealpha amylase is an acid stable alpha amylase. In one embodiment, thealpha amylase is a granular starch hydrolyzing enzyme (GSHE). In oneembodiment, the alpha amylase is Aspergillus kawachi alpha amylase(AKAA), see U.S. Pat. No. 7,037,704. Commercially available alphaamylases contemplated for use in the compositions of the disclosure areknown and include GZYME G997, SPEZYME FRED, SPEZYME XTRA, STARGEN(Danisco US, Inc, Genencor Division), TERMAMYL 120-L and SUPRA(Novozymes, Biotech.) and VIRIDIUM (Diversa).

In other embodiments, the variant glucoamylases of the disclosure may becombined with other glucoamylases. In some embodiments, theglucoamylases of the disclosure will be combined with one or moreglucoamylases derived from strains of Aspergillus or variants thereof,such as A. oryzae, A. niger, A. kawachi, and A. awamori; glucoamylasesderived from strains of Humicola or variants thereof, particularly H.grisea, such as the glucoamylase having at least 90%, 93%, 95%, 96%,97%, 98% and 99% sequence identity to SEQ ID NO: 3 disclosed in WO05/052148; glucoamylases derived from strains of Talaromyces or variantsthereof, particularly T. emersonii; glucoamylases derived from strainsof Athelia and particularly A. rolfsii; glucoamylases derived fromstrains of Penicillium, particularly P. chrysogenum.

14. USES

In particular, the variant glucoamylases may be used for starchconversion processes, and particularly in the production of dextrose forfructose syrups, specialty sugars and in alcohol and other end-product(e.g., organic acid, ascorbic acid, and amino acids) production fromfermentation of starch containing substrates (G. M. A van Beynum et al.,Eds. (1985) STARCH CONVERSION TECHNOLOGY, Marcel Dekker Inc. NY).Dextrins produced using variant glucoamylase compositions of thedisclosure may result in glucose yields of at least 80%, at least 85%,at least 90% and at least 95%. Production of alcohol from thefermentation of starch substrates using glucoamylases encompassed by thedisclosure may include the production of fuel alcohol or portablealcohol. In some embodiments, the production of alcohol will be greaterwhen the variant glucoamylase is used under the same conditions as theparent glucoamylase. In some embodiments, the production of alcohol willbe between about 0.5% and 2.5% better, including but not limited to0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%, 1.6%. 1.7%,1.8%, 1.9%, 2.0%, 2.1%, 2.2%, 2.3%, and 2.4% more alcohol than theparent glucoamylase.

In one representative embodiment, the variant glucoamylases of thedisclosure will find use in the hydrolysis of starch from variousplant-based substrates, which are used for alcohol production. In someembodiments, the plant-based substrates will include corn, wheat,barley, rye, milo, rice, sugar cane, potatoes and combinations thereof.In some embodiments, the plant-based substrate will be fractionatedplant material, for example a cereal grain such as corn, which isfractionated into components such as fiber, germ, protein and starch(endosperm) (U.S. Pat. No. 6,254,914 and U.S. Pat. No. 6,899,910).Methods of alcohol fermentations are described in THE ALCOHOL TEXTBOOK,A REFERENCE FOR THE BEVERAGE, FUEL AND INDUSTRIAL ALCOHOL INDUSTRIES,3^(rd) Ed., Eds K. A. Jacques et al., 1999, Nottingham University Press,UK. In certain embodiments, the alcohol will be ethanol. In particular,alcohol fermentation production processes are characterized as wetmilling or dry milling processes. In some embodiments, the variantglucoamylase will be used in a wet milling fermentation process and inother embodiments the variant glucoamylase will find use in a drymilling process.

Dry grain milling involves a number of basic steps, which generallyinclude: grinding, cooking, liquefaction, saccharification, fermentationand separation of liquid and solids to produce alcohol and otherco-products. Plant material and particularly whole cereal grains, suchas corn, wheat or rye are ground. In some cases the grain may be firstfractionated into component parts. The ground plant material may bemilled to obtain a coarse or fine particle. The ground plant material ismixed with liquid (e.g., water and/or thin stillage) in a slurry tank.The slurry is subjected to high temperatures (e.g., 90° C. to 105° C. orhigher) in a jet cooker along with liquefying enzymes (e.g., alphaamylases) to solubilize and hydrolyze the starch in the grain todextrins. The mixture is cooled down and further treated withsaccharifying enzymes, such as glucoamylases encompassed by the instantdisclosure, to produce glucose. The mash containing glucose may then befermented for approximately 24 to 120 hours in the presence offermentation microorganisms, such as ethanol producing microorganism andparticularly yeast (Saccharomyces spp.). The solids in the mash areseparated from the liquid phase and alcohol such as ethanol and usefulco-products such as distillers' grains are obtained.

In some embodiments, the saccharification step and fermentation step arecombined and the process is referred to as simultaneous saccharificationand fermentation or simultaneous saccharification, yeast propagation andfermentation.

In other embodiments, the variant glucoamylase is used in a process forstarch hydrolysis wherein the temperature of the process is carried outat a temperature of between 30° C. and 75° C. and also at a temperatureof between 40° C. and 65° C. at a pH range of between pH 3.0 and pH 6.5.The fermentation processes in some embodiments include milling of acereal grain or fractionated grain and combining the ground cereal grainwith liquid to form a slurry that is then mixed in a single vessel witha variant glucoamylase according to the disclosure and optionally otherenzymes such as but not limited to alpha amylases, other glucoamylases,phytases, proteases, pullulanases, isoamylases or other enzymes havinggranular starch hydrolyzing activity and yeast to produce ethanol andother co-products (U.S. Pat. No. 4,514,496, WO 04/081193 and WO04/080923).

In some embodiments, the disclosure pertains to a method ofsaccharifying a liquid starch solution, which comprises an enzymaticsaccharification step using a variant glucoamylase of the disclosure.

The present disclosure also provides an animal feed comprising at leastone variant glucoamylase encompassed by the disclosure. Methods of usinga glucoamylase enzyme in the production of feeds comprising starch areprovided in WO 03/049550, filed Dec. 13, 2002 (herein incorporated byreference in its entirety). Briefly, the glucoamylase variant is admixedwith a feed comprising starch. The glucoamylase is capable of degradingresistant starch for use by the animal.

Other objects and advantages of the present disclosure are apparent fromthe present Specification.

EXAMPLES

The following examples are provided in order to demonstrate and furtherillustrate certain representative embodiments and aspect of the presentdisclosure and are not to be construed as limiting the scope thereof.

In the disclosure and experimental section that follows, the followingabbreviations apply: GA (glucoamylase); GAU (glucoamylase unit); wt %(weight percent); ° C. (degrees Centigrade); rpm (revolutions perminute); H₂O (water); dH₂O (deionized water); dIH₂O (deionized water,Milli-Q filtration); aa or AA (amino acid); bp (base pair); kb (kilobasepair); kD (kilodaltons); g or gm (grams); μg (micrograms); mg(milligrams); μL (microliters); ml and mL (milliliters); mm(millimeters); μm (micrometer); M (molar); mM (millimolar); μM(micromolar); U (units); V (volts); MW (molecular weight); sec(s) ors(s) (second/seconds); min(s) or m(s) (minute/minutes); hr(s) or h(s)(hour/hours); DO (dissolved oxygen); ABS (Absorbance); EtOH (ethanol);PSS (physiological salt solution; m/v (mass/volume); and MTP (microtiterplate); N (Normal); DP1 (monosaccharides); DP2 (disaccharides); DP>3(oligosaccharides, sugars having a degree of polymerization greater than3); ppm (parts per million).

The methods used to provide variants are described below. However, itshould be noted that different methods may be used to provide variantsof a parent molecule and the disclosure is not limited to the methodsused in the examples. It is intended that any suitable means for makingvariants and selection of variants may be used.

pNPG Glucoamylase Activity Assay for 96-Well Microtiter Plates:

The reagent solutions were: NaAc buffer: 200 mM sodium acetate buffer pH4.5; Substrate: 50 mM p-nitrophenyl-α-D-glucopyranoside (Sigma N-1377)in NaAc buffer (0.3 g/20 ml) and stop solution: 800 mM glycine-NaOHbuffer pH 10. 30 μl filtered supernatant was placed in a fresh 96-wellflat bottom MTP. To each well 50 μl NaAc buffer and 120 μl substrate wasadded and incubated for 30 minutes at 50° C. (Thermolab systems iEMSIncubator/shaker HT). The reaction was terminated by adding 100 μl stopsolution. The absorbance was measured at 405 nm in a MTP-reader(Molecular Devices Spectramax 384 plus) and the activity was calculatedusing a molar extinction coefficient of 0.011 μM/cm.

Thermal Stability Assay:

Crude supernatant (100 μl) was added to 100 μl 50 mM NaAc buffer pH 4.5.the sample was equally divided over 2 MTP. One MTP (initial plate) wasincubated for 1 hr at 4° C. and the other MTP (residual plate) wasincubated at 60° C. (Thermolab systems iEMS Incubator/Shaker HT) for 1hr. The residual plate was chilled for 15 min on ice. Activity ismeasured on both plates using the ethanol application assay describedbelow, with the following modification: the amount of sample taken forthe thermostability assay is 25 μl and the amount of 30 mM NaAc bufferpH 4.0 is 35 μl.

Thermostability is calculated as % residual activity as follows:

$\frac{{{ABS}(340)}{residual}\text{-}{blank}}{{{ABS}(340)}{initial}\text{-}{blank}} \times 100\%$

The crude supernatant material is tested for remaining glucose in theculture medium after the growth period. If remaining glucose is found,the absorbance value is subtracted from the measured absorbance valuesof both the initial activity as the residual activity.

Bradford Assay for Protein Quantification in 96-Well Microtiter Plates:

The reagent solution was Bradford Quickstart work solution (BioRadcat#500-0205). 100 μl of 10 kD-filtered supernatant was placed in afresh 96-well flat bottom plate. To each well 200 μl reagent was addedand incubated for 5 minutes at room temperature. The absorbance wasmeasured at 595 nm in a MTP-reader (Molecular Devices Spectramax 384plus). Protein concentrations were calculated according to a BovineSerum Albumin (BSA) (0-50 μg/ml) standard curve.

Hexokinase Activity Assay:

Hexokinase cocktail: 10-15 minutes prior to use, 90 ml water was addedto a BoatIL container glucose HK R1 (IL test glucose (HK) kit,Instrument Laboratory #182507-40) and gently mixed. 85 μl of Hexokinasecocktail were added to 100 μl of dH₂O. 15 μl of sample were added to themixtures and incubated for 5 mins in the dark at room temperature.Absorbance was read at 340 nm in a MTP-reader. Glucose concentrationswere calculated according to a glucose (0-1 mg/ml) standard curve.

Assay Conditions Ethanol Application:

To prepare the 8% stock solution, 8 g of soluble corn starch (Sigma#S4180) was suspended in 40 ml dH₂O at room temperature. Fiftymilliliters of boiling dH₂O was added to the slurry in a 250 ml flaskand cooked for 5 minutes. The starch solution was cooled to 25° C. andthe volume adjusted to 100 ml with dH₂O. The 4% (m/v) soluble starchworking solution was prepared by diluting (1:1) the stock solution with100 mM sodium acetate buffer pH 3.7.

For the screening assay, 5 μl crude supernatant was diluted with 175 μl50 mM NaAc buffer pH 4.5 in a flat bottom 96-well MTP. Sixty microlitersof this dilution was added to 120 μl 4% soluble corn starch andincubated at 900 rpm for 2 hrs at 32° C. (Thermolab systems iEMSIncubator/Shaker HT). The reaction was stopped by adding 90 μl 4°C.-cold Stop Solution (800 mM Glycine-NaOH buffer, pH 10). The samplewas placed on ice. Starch was spun down at 1118×g at 15° C. for 5minutes (SIGMA 6K15) and 15 μl supernatant was used in the Hexokinaseactivity assay described above to determine the glucose content.

The crude supernatant material was tested for remaining glucose in theculture medium after the growth period. If remaining glucose was found,the amount of glucose produced by the glucoamylase was not calculated.

Assay Conditions Sweetener Application:

To prepare the 8% stock solution, 8 g of soluble starch (Sigma #S4180)was suspended in 40 ml water at room temperature. Then, 50 ml of boilingdH₂O was added to the slurry in a 250 ml flask and cooked for 5 mins.The starch solution was cooled to 25° C. and the volume adjusted to 100ml with dH₂O. The 4% (m/v) soluble starch working solution was preparedby diluting stock solution 1:1 with 100 mM sodium acetate buffer pH 4.5.

Fifty microliters of 80 mM NaAc buffer pH 4.5 were placed in a fresh96-well flat bottom plate. To each well, 120 μl 4% soluble cornstarchand 5 μl 10 kD-filtered supernatant was added and incubated for 1 hr at60° C. The reaction was stopped by adding 90 μl 4° C. cold Stop Solution(800 mM Glycine-NaOH buffer, pH 10.0). The sample was placed on ice for30 mins. Starch was spun down at 716 rpm at 15° C. for 5 minutes (Sigma6K15, centrifuge) and 15 μl of the supernatant was used in theHexokinase activity assay described herein to determine glucose content.

Example 1 Construction of the pREP3Y-TrGA Vector

The TrGA expression cassette composed of the DNA sequence (SEQ ID NO: 4)encoding the TrGA signal peptide, the pro-sequence, and the matureprotein, including the catalytic domain, linker region and starchbinding domain, was cloned into pDONR™201, a Gateways Entry vector(Invitrogen, Carlsbad, Calif., USA). The TrGA expression cassette wascloned into the Gateway compatible destination vector pREP3Y-DEST (FIG.3) by the Gateways LR recombination reaction.

The pRep3Y-TrGA expression vector (FIG. 3B) enabled the expression ofthe TrGA protein (SEQ ID NO: 2) in Schizosaccharomyces pombe.

Sixty-five TrGA site saturated mutagenesis (SSM) libraries wereconstructed using the pDONR-TrGA entry vector (FIG. 2) as a template andthe primers listed in Table 2. The mutagenesis primers used in theexperiments all contain the triple DNA sequence code NNS (N=A,C,T,G andS═C or G) at the position that corresponds with the codon of the TrGAsequence to be mutated (SEQ ID NO: 2) and initiated random incorporationof nucleotides at that position. Construction of each SSM librarystarted with two PCR amplifications using the Gateway forward(pDONR201-FW) and a specific reverse mutagenesis primer (Table 3), andGateway reversed (pDONR201-RV) primer and a specific forward mutagenesisprimer (Table 2) (equal positions for the mutagenesis primers). Phusionhigh fidelity DNA polymerase (Finnzymes OY, Espoo, Finland) was used forPCR amplification (0.2 μM primers, 25 cycles) according to the protocolprovided by Finnzymes. Briefly, 1 μl (SEQ ID NO: 1), DNA fragment ofboth specific PCR mixes, both targeted at the same codon were added to48 μl of fresh PCR reaction solution together with primers Gateway FWand Gateway RV (Invitrogen) and mixed. This fusion PCR amplification (22cycles) resulted in a linear expression cassette DNA fragment with aspecific TrGA codon randomly mutated and unique Gateway recombinationsites on both ends. Purification of this DNA fragment (ChargeSwitch® PCRclean-up, Invitrogen, Carlsbad USA) and a BP recombination reaction(Invitrogen, Carlsbad, USA) with pDONR201 (Invitrogen) generated acircular multimeric DNA (entry vector) that was subsequently transformedto E. coli Max efficiency DH5α (Invitrogen) and plated on 2×TY medium[Bacto Tryptone (Difco) 16 g/L, Bacto Yeast Extract (Difco) 10 g/L, NaCL5 g/L] supplemented with 50 μg/mL kanamycin.

TABLE 2 Forward primers used to generate TrGA SSM libraries SEQ IDPrimers DNA SEQUENCE 5′ TO 3′ NO: pDONR201- TCGCGTTAACGCTAGCATGGATCTC 10FW D4F TCTGTTGACNNSTTCATCAGCACCGAGACGC 11 F5FTCTGTTGACGACNNSATCAGCACCGAGACGCCTA 12 I12FATCAGCACCGAGACGCCTNNSGCACTGAACAATCTTCTTT 13 D24FCTTTGCAATGTTGGTCCTNNSGGATGCCGTGCATTCGGCA 14 F29FCCTGATGGATGCCGTGCANNSGGCACATCAGCTGGTGCGG 15 I43FATTGCATCTCCCAGCACANNSGACCCGGACTACTATTACA 16 D44FGCATCTCCCAGCACAATTNNSCCGGACTACTATTACATGT 17 P45FTCTCCCAGCACAATTGACNNSGACTACTATTACATGTGGA 18 D46FCCCAGCACAATTGACCCGNNSTACTATTACATGTGGACGC 19 Y47FAGCACAATTGACCCGGACNNSTATTACATGTGGACGCGAG 20 Y49FATTGACCCGGACTACTATNNSATGTGGACGCGAGATAGCG 21 W51FCCGGACTACTATTACATGNNSACGCGAGATAGCGCTCTTG 22 Y70FGACCGCTTCACCGAAACGNNSGATGCGGGCCTGCAGCGCC 23 Q75FACGTACGATGCGGGCCTGNNSCGCCGCATCGAGCAGTACA 24 R76FTACGATGCGGGCCTGCAGNNSCGCATCGAGCAGTACATTA 25 P94FCTCCAGGGCCTCTCTAACNNSTCGGGCTCCCTCGCGGACG 26 D100FCCCTCGGGCTCCCTCGCGNNSGGCTCTGGTCTCGGCGAGC 27 K114FAAGTTTGAGTTGACCCTGNNSCCTTTCACCGGCAACTGGG 28 F116FGAGTTGACCCTGAAGCCTNNSACCGGCAACTGGGGTCGAC 29 N119FCTGAAGCCTTTCACCGGCNNSTGGGGTCGACCGCAGCGGG 30 R122FTTCACCGGCAACTGGGGTNNSCCGCAGCGGGATGGCCCAG 31 R125FAACTGGGGTCGACCGCAGNNSGATGGCCCAGCTCTGCGAG 32 N146FAAGTGGCTCATCAACAACNNSTATCAGTCGACTGTGTCCA 33 Q148FCTCATCAACAACAACTATNNSTCGACTGTGTCCAACGTCA 34 Y169FCTCAACTATGTTGCCCAGNNSTGGAACCAAACCGGCTTTG 35 Q172FGTTGCCCAGTACTGGAACNNSACCGGCTTTGACCTCTGGG 36 F175FTACTGGAACCAAACCGGCNNSGACCTCTGGGAAGAAGTCA 37 W178FCAAACCGGCTTTGACCTCNNSGAAGAAGTCAATGGGAGCT 38 E180FGGCTTTGACCTCTGGGAANNSGTCAATGGGAGCTCATTCT 39 V181FTTTGACCTCTGGGAAGAANNSAATGGGAGCTCATTCTTTA 40 Q208FCTTGCTGCCACTCTTGGCNNSTCGGGAAGCGCTTATTCAT 41 S211FACTCTTGGCCAGTCGGGANNSGCTTATTCATCTGTTGCTC 42 W228FTGCTTTCTCCAACGATTCNNSGTGTCGTCTGGTGGATACG 43 N242FGACTCCAACATCAACACCNNSGAGGGCAGGACTGGCAAGG 44 E243FTCCAACATCAACACCAACNNSGGCAGGACTGGCAAGGATG 45 R245FATCAACACCAACGAGGGCNNSACTGGCAAGGATGTCAACT 46 I292FGTCGACTCCTTCCGCTCCNNSTACGGCGTGAACAAGGGCA 47 G294FTCCTTCCGCTCCATCTACNNSGTGAACAAGGGCATTCCTG 48 K297FTCCATCTACGGCGTGAACNNSGGCATTCCTGCCGGTGCTG 49 R309FGCTGCCGTCGCCATTGGCNNSTATGCAGAGGATGTGTACT 50 Y310FGCCGTCGCCATTGGCCGGNNSGCAGAGGATGTGTACTACA 51 D313FATTGGCCGGTATGCAGAGNNSGTGTACTACAACGGCAACC 52 V314FGGCCGGTATGCAGAGGATNNSTACTACAACGGCAACCCTT 53 Y315FCGGTATGCAGAGGATGTGNNSTACAACGGCAACCCTTGGT 54 Y316FTATGCAGAGGATGTGTACNNSAACGGCAACCCTTGGTATC 55 N317FGCAGAGGATGTGTACTACNNSGGCAACCCTTGGTATCTTG 56 W321FTACTACAACGGCAACCCTNNSTATCTTGCTACATTTGCTG 57 K340FGATGCCATCTACGTCTGGNNSAAGACGGGCTCCATCACGG 58 K341FGCCATCTACGTCTGGAAGNNSACGGGCTCCATCACGGTGA 59 T350FTCCATCACGGTGACCGCCNNSTCCCTGGCCTTCTTCCAGG 60 Q356FACCTCCCTGGCCTTCTTCNNSGAGCTTGTTCCTGGCGTGA 61 T363FGAGCTTGTTCCTGGCGTGNNSGCCGGGACCTACTCCAGCA 62 S368FGTGACGGCCGGGACCTACNNSAGCAGCTCTTCGACCTTTA 63 S369FACGGCCGGGACCTACTCCNNSAGCTCTTCGACCTTTACCA 64 N376FAGCTCTTCGACCTTTACCNNSATCATCAACGCCGTCTCGA 65 Y395FCTCAGCGAGGCTGCCAAGNNSGTCCCCGCCGACGGTTCGC 66 A398FGCTGCCAAGTACGTCCCCNNSGACGGTTCGCTGGCCGAGC 67 S401FTACGTCCCCGCCGACGGTNNSCTGGCCGAGCAGTTTGACC 68 R408FCTGGCCGAGCAGTTTGACNNSAACAGCGGCACTCCGCTGT 69 N409FGCCGAGCAGTTTGACCGCNNSAGCGGCACTCCGCTGTCTG 70 T412FTTTGACCGCAACAGCGGCNNSCCGCTGTCTGCGCTTCACC 71 H418FACTCCGCTGTCTGCGCTTNNSCTGACGTGGTCGTACGCCT 72 W421FTCTGCGCTTCACCTGACGNNSTCGTACGCCTCGTTCTTGA 73 R433FTTGACAGCCACGGCCCGTNNSGCTGGCATCGTGCCCCCCT 74 I436FACGGCCCGTCGGGCTGGCNNSGTGCCCCCCTCGTGGGCCA 75 S451FAGCGCTAGCACGATCCCCNNSACGTGCTCCGGCGCGTCCG 76

TABLE 3 Primers used to generate TrGA SSM libraries SEQ ID Primer DNAsequence 5′ to 3′ NO: pDON201- GTAACATCAGAGATTTTGAGACAC 77 RV D4RGCGTCTCGGTGCTGATGAASNNGTCAACAGA 78 F5RTAGGCGTCTCGGTGCTGATSNNGTCGTCAACAGA 79 I12RAAAGAAGATTGTTCAGTGCSNNAGGCGTCTCGGTGCTGAT 80 D24RTGCCGAATGCACGGCATCCSNNAGGACCAACATTGCAAAG 81 F29RCCGCACCAGCTGATGTGCCSNNTGCACGGCATCCATCAGG 82 I43RTGTAATAGTAGTCCGGGTCSNNTGTGCTGGGAGATGCAAT 83 D44RACATGTAATAGTAGTCCGGSNNAATTGTGCTGGGAGATGC 84 P45RTCCACATGTAATAGTAGTCSNNGTCAATTGTGCTGGGAGA 85 D46RGCGTCCACATGTAATAGTASNNCGGGTCAATTGTGCTGGG 86 Y47RCTCGCGTCCACATGTAATASNNGTCCGGGTCAATTGTGCT 87 Y49RCGCTATCTCGCGTCCACATSNNATAGTAGTCCGGGTCAAT 88 W51RCAAGAGCGCTATCTCGCGTSNNCATGTAATAGTAGTCCGG 89 Y70RGGCGCTGCAGGCCCGCATCSNNCGTTTCGGTGAAGCGGTC 90 Q75RTGTACTGCTCGATGCGGCGSNNCAGGCCCGCATCGTACGT 91 R76RTAATGTACTGCTCGATGCGSNNCTGCAGGCCCGCATCGTA 92 P94RCGTCCGCGAGGGAGCCCGASNNGTTAGAGAGGCCCTGGAG 93 D100RGCTCGCCGAGACCAGAGCCSNNCGCGAGGGAGCCCGAGGG 94 K114RCCCAGTTGCCGGTGAAAGGSNNCAGGGTCAACTCAAACTT 95 F116RGTCGACCCCAGTTGCCGGTSNNAGGCTTCAGGGTCAACTC 96 N119RCCCGCTGCGGTCGACCCCASNNGCCGGTGAAAGGCTTCAG 97 R122RCTGGGCCATCCCGCTGCGGSNNACCCCAGTTGCCGGTGAA 98 R125RCTCGCAGAGCTGGGCCATCSNNCTGCGGTCGACCCCAGTT 99 N146RTGGACACAGTCGACTGATASNNGTTGTTGATGAGCCACTT 100 Q148RTGACGTTGGACACAGTCGASNNATAGTTGTTGTTGATGAG 101 Y169RCAAAGCCGGTTTGGTTCCASNNCTGGGCAACATAGTTGAG 102 Q172RCCCAGAGGTCAAAGCCGGTSNNGTTCCAGTACTGGGCAAC 103 F175RTGACTTCTTCCCAGAGGTCSNNGCCGGTTTGGTTCCAGTA 104 W178RAGCTCCCATTGACTTCTTCSNNGAGGTCAAAGCCGGTTTG 105 E180RAGAATGAGCTCCCATTGACSNNTTCCCAGAGGTCAAAGCC 106 V181RTAAAGAATGAGCTCCCATTSNNTTCTTCCCAGAGGTCAAA 107 Q208RATGAATAAGCGCTTCCCGASNNGCCAAGAGTGGCAGCAAG 108 S211RGAGCAACAGATGAATAAGCSNNTCCCGACTGGCCAAGAGT 109 W228RCGTATCCACCAGACGACACSNNGAATCGTTGGAGAAAGCA 110 N242RCCTTGCCAGTCCTGCCCTCSNNGGTGTTGATGTTGGAGTC 111 E243RCATCCTTGCCAGTCCTGCCSNNGTTGGTGTTGATGTTGGA 112 R245RAGTTGACATCCTTGCCAGTSNNGCCCTCGTTGGTGTTGAT 113 I292RTGCCCTTGTTCACGCCGTASNNGGAGCGGAAGGAGTCGAC 114 G294RCAGGAATGCCCTTGTTCACSNNGTAGATGGAGCGGAAGGA 115 K297RCAGCACCGGCAGGAATGCCSNNGTTCACGCCGTAGATGGA 116 R309RAGTACACATCCTCTGCATASNNGCCAATGGCGACGGCAGC 117 Y310RTGTAGTACACATCCTCTGCSNNCCGGCCAATGGCGACGGC 118 D313RGGTTGCCGTTGTAGTACACSNNCTCTGCATACCGGCCAAT 119 V314RAAGGGTTGCCGTTGTAGTASNNATCCTCTGCATACCGGCC 120 Y315RACCAAGGGTTGCCGTTGTASNNCACATCCTCTGCATACCG 121 Y316RGATACCAAGGGTTGCCGTTSNNGTACACATCCTCTGCATA 122 N317RCAAGATACCAAGGGTTGCCSNNGTAGTACACATCCTCTGC 123 W321RCAGCAAATGTAGCAAGATASNNAGGGTTGCCGTTGTAGTA 124 K340RCCGTGATGGAGCCCGTCTTSNNCCAGACGTAGATGGCATC 125 K341RTCACCGTGATGGAGCCCGTSNNCTTCCAGACGTAGATGGC 126 T350RCCTGGAAGAAGGCCAGGGASNNGGCGGTCACCGTGATGGA 127 Q356RTCACGCCAGGAACAAGCTCSNNGAAGAAGGCCAGGGAGGT 128 T363RTGCTGGAGTAGGTCCCGGCSNNCACGCCAGGAACAAGCTC 129 S368RTAAAGGTCGAAGAGCTGCTSNNGTAGGTCCCGGCCGTCAC 130 S369RTGGTAAAGGTCGAAGAGCTSNNGGAGTAGGTCCCGGCCGT 131 N376RTCGAGACGGCGTTGATGATSNNGGTAAAGGTCGAAGAGCT 132 Y395RGCGAACCGTCGGCGGGGACSNNCTTGGCAGCCTCGCTGAG 133 A398RGCTCGGCCAGCGAACCGTCSNNGGGGACGTACTTGGCAGC 134 S401RGGTCAAACTGCTCGGCCAGSNNACCGTCGGCGGGGACGTA 135 R408RACAGCGGAGTGCCGCTGTTSNNGTCAAACTGCTCGGCCAG 136 N409RCAGACAGCGGAGTGCCGCTSNNGCGGTCAAACTGCTCGGC 137 T412RGGTGAAGCGCAGACAGCGGSNNGCCGCTGTTGCGGTCAAA 138 H418RAGGCGTACGACCACGTCAGSNNAAGCGCAGACAGCGGAGT 139 W421RTCAAGAACGAGGCGTACGASNNCGTCAGGTGAAGCGCAGA 140 R433RAGGGGGGCACGATGCCAGCSNNACGGGCCGTGGCTGTCAA 141 I436RTGGCCCACGAGGGGGGCACSNNGCCAGCCCGACGGGCCGT 142 S451RCGGACGCGCCGGAGCACGTSNNGGGGATCGTGCTAGCGCT 143

For each library, after overnight incubation at 37° C., colonies werepooled by resuspension of the clones in PSS. From the pooled E. colitransformants, total plasmid was isolated (Qiagen) using standardtechniques. Briefly 1 μl of the plasmid solution was added to 1 μl ofpRep3Y destination vector (FIG. 1A) solution and added to the LRCLONASE™ II enzyme mix according to the protocol supplied by Invitrogen.A circular multimeric DNA was generated and transformed to E. coli MaxEfficiencyDH5α as described by the supplier.

After overnight incubation at 37° C., 96 single colonies of each librarywere picked from 2×TY agar plates with 100 μg/ml ampicillin and grownfor 24 hrs at 37° C. in a MTP containing 200 μL 2×TY medium with 100μg/ml ampicillin. Cultures were used for sequence analyses (BaseClearB.V., Leiden, Netherlands).

The library numbers ranged from 1 to 65 with an addition referring tothe codon of the TrGA sequence that is randomly mutated. Afterselection, each library included a maximum of 19 TrGA variants. Thesevariants were individually transferred to Schizosaccharomyces pombeaccording to manufacturers instruction. (Zymo Research, Orange Calif.USA).

S. pombe transformations were plated on selective medium (EMM agar,Qbiogene, Irvine, USA Cat. No. 4110-232) and incubated at 28° C. for 4days. Transformants were purified from the transformation plate bystreaking the colonies on EMM agar.

Example 2 Description of the Growth Conditions and the SamplePre-Treatment

S. pombe transformants were inoculated in 96 well microtiter plates(MTP) containing selective medium (2×EMM-C) [64.4 g/L EMM Broth(Qbiogene Cat. No: 4110-032), 0.62 g/L Complete Supplement Mixture(CSM-HIS-LEU-TRP, Qbiogene, Cat. No. 4530-122)] and incubated overnightat 28° C. From the overnight incubated microtiter plate, 200 μl of grownS. pombe culture was inoculated in 20 ml of 2×EMM-C liquid medium in a100 ml shake flask and incubated for 4 days at 26° C. at 280 rpm in aMultitron shaking incubator (Infors AG, Bottmingen, Switzerland). Fromthe grown culture, 2 ml of S. pombe culture was sampled and centrifugedfor 5 min at 14,000 rpm (Sigma). The supernatant was transferred into a10 kD Vivaspin 500 HT filter set-up (VivaScience AG, Hannover, Germany)and centrifuged for 25 min at 1000 g. The retentate was diluted back tothe original start volume with 50 mM NaAc pH 4.5 supplemented with0.015% Tween-80. This solution was used in the different assays.

Example 3 Construction of a Combinatorial Library of 4 Variants of TrGA

(A) Experiments were conducted for the construction of TrGA-variantscarrying combinations of the following single site mutations: Q172F;Q208N; S211R and V314H. A review of the variants is shown below:

a) Q172F; Q208N

b) Q172F; S211R

c) Q172F; V314H

d) Q208N; S211R

e) Q208N V314H

f) S211R; V314H

g) Q172F; Q208N; S211R

h) Q172F; Q208N; V314H

i) Q172F; S211R; V314H

j) Q208N; S211R; V314H

k) Q172F; Q208N; S211R; V314H

The Quikchange® multi site-directed mutagenesis (QCMS) kit (Stratagene)was used to construct the library. The 5′ phosphorylated primers used tocreate the library are shown in Table 4. Optimal results in terms ofincorporation of full length primers as well as significant reduction inprimer-derived errors were obtained by the use of HPLC, PAGE or anyother type of purified primers (Invitrogen).

TABLE 4 Primers used to construct selected combinatorial variants SEQ IDPrimer Sequence NO: I43R GCATCTCCCAGCACACGAGACCCGGACTACTAT 144 I43YGCATCTCCCAGCACATACGACCCGGACTACTAT 145 R76LGATGCGGGCCTGCAGCTGCGCATCGAGCAGTAC 146 N119TCTGAAGCCTTTCACCGGCACCTGGGGTCGACCGCAGCGGG 147 N119YTGAAGCCTTTCACCGGCTACTGGGGTCGACCGCAGCGGG 148 N119DCTGAAGCCTTTCACCGGCGACTGGGGTCGACCGCAGCGGG 149 N146AGTGGCTCATCAACAACGASTATCAGTCGACTGTGT 150 N146TAGTGGCTCATCAACAACACCTATCAGTCGACTGTGT 151 N146WGTGGCTCATCAACAATGGTATCAGTCGACTGTGT 152 N146LAGTGGCTCATCAACAACCTGTATCAGTCGACTGTGT 153 N146SAGTGGCTCATCAACAACTCCTATCAGTCGACTGTGT 154 Q172D/TTGCCCAGTACTGGAACGASACCGGCTTTGACCTCTGG 155 E Q172V/TTGCCCAGTACTGGAACSTGACCGGCTTTGACCTCTGG 156 L Q172TTTGCCCAGTACTGGAACACCACCGGCTTTGACCTCTGG 157 Q172RTTGCCCAGTACTGGAACCGAACCGGCTTTGACCTCTGG 158 Q172CTTGCCCAGTACTGGAACTGCACCGGCTTTGACCTCTGG 159

The template plasmid pDONR-TrGA (FIG. 2) was used to construct thecombinatorial library using the QCMS kit of Stratagene. The library wasconstructed as described by the supplier with modified primerconcentrations used in the reactions. Specifically, 4 μl pDONR-TrGA(25-50 ng) was mixed with 11 μl of sterile distilled water; 1.5 μl ofdNTP; 2.5 μl of 10×QCMS-buffer; 1 μl of enzyme blend and 1 μl of eachmutant primer mix giving a total of 100 ng of primers in each reaction.The PCR conditions were 95° C. for 1 min, followed by 30 cycles of 95°for 1 min, 55° C. for 1 min, and 65° C. for 6 min, in an MJ Researchthermocycler using thin-walled 0.2 ml PCR tubes. The reaction productwas digested with 1 μl of Dpn1 from the QCMS kit by incubating at 37° C.overnight. A PCR purification kit (Qiagen) was used for samplepurification and a second round of digestion was carried out with Dpn1(Stratagene) for 1 hour at 37° C.

The reaction mixture was transformed to into E. coli Max efficiency DH5α(Invitrogen) and plated on selective agar (2×TY supplemented with 50 μgkanamycin/ml). After overnight incubation at 37° C., 96 single colonieswere picked for sequence analysis (BaseClear B.V., Leiden, Netherlands).The combinatorial variants were cloned and expressed in a T. reesei hoststrain as described below and in WO 06/060062.

(B) A further six combinatorial libraries (Table 5) were syntheticallymade by Geneart (Regensburg, Germany) and were tested for thermalstability and in ethanol and sweetener application assays as describedherein.

TABLE 5 Combinatorial libraries 1 D24E, L/I43F, R/D44H, N/F175H/V181K,L/V314D, H, K/T363R 2 D24L, W, Y/Q208F/I292F, L, N, V/G294A, I,Q/K297A/Y310F, Q, R 3 V181F, K, L/E243A, N, M, R, Y/I292F, L, N,V/K297A, D, H, M, N, Q/N317H/Y395Q, R 4 D24E, L, Y/V181F, K,L/Q208C/F/G294A, I, Q/T363R/N376Q/ N409K, W 5 D24E, L, Y/V181F, K,L/I292F, L, N, V/G294A, I, Q/E243A, M, N, R, Y/N409K, W 6 I43R/E243A, M,N, R, Y/I292F, L, N, V/G294A/ K297A, D, H, M, N, Q, S, R, W, Y

Example 4 Variants with Improved Thermal Stability

The parent TrGA molecule under the conditions described had a residualactivity between 15 and 44% (day-to-day variation). The performanceindex was calculated based on the TrGA thermostability of the samebatch. The performance indices are the quotients PI=(Variant residualactivity)/(TrGA residual activity). A performance index >1 indicates animproved stability. Variants that have a thermal stability performanceindex of more than 1.0 are shown in the following Table 6.

TABLE 6 Thermostability screening PI Thermal Stability, 60° C., VariantpH 4.5 D4P 1.05 I12E 1.09 I12Y 1.40 D24L 1.09 D24W 1.13 D24Y 1.03 I43R1.28 D44N 1.06 D44Q 1.10 Q75N 1.09 R76K 1.03 N146D 1.20 N146E 1.24 N146L1.10 N146V 1.28 N146W 1.17 Q148D 1.02 F175I 1.02 F175Y 1.06 E180A 1.41E180D 1.02 E180G 1.13 E180I 1.41 E180L 1.38 E180M 1.10 E180N 1.27 E180Q1.72 E180R 1.59 E180V 1.08 E180W 1.30 E180Y 1.31 V181I 1.20 V181K 1.12V181L 1.06 V181Q 1.09 V181R 1.07 Q208F 1.06 Q208T 1.17 Q208V 1.15 S211D1.10 S211E 1.02 S211I 1.31 S211M 1.90 E243A 1.19 E243H 1.04 E243M 1.53E243N 1.35 E243P 1.06 E243R 1.21 E243S 1.09 E243T 1.48 E243Y 1.43 I292F1.17 I292L 1.10 I292N 1.31 I292V 1.02 G294A 1.30 G294C 1.41 G294D 1.31G294E 1.34 G294H 1.17 G294I 2.15 G294L 2.01 G294P 1.13 G294Q 1.91 G294R1.34 G294V 1.10 K297A 1.47 K297C 1.10 K297D 1.50 K297F 1.24 K297G 1.25K297H 1.63 K297L 1.62 K297M 1.62 K297N 1.87 K297Q 1.82 K297R 1.29 K297S1.22 K297T 1.33 K297V 1.10 K297W 1.85 K297Y 1.71 R309S 1.08 Y310C 1.06Y310F 1.35 Y310L 1.11 Y310Q 1.40 Y310R 1.61 Y315E 1.24 Y315H 1.48 Y315L1.35 Y315N 1.17 Y315P 1.19 Y315Q 1.43 Y315T 1.34 Y316D 1.06 N317H 1.26N317Q 1.09 K340H 1.02 K340R 1.09 K341I 1.10 K341V 1.07 T350G 1.08 T350P1.08 T350S 1.33 Q356L 1.20 T363N 1.30 S368C 1.12 S368E 1.07 S368F 1.16S368H 1.26 S368I 1.15 S368L 1.33 S368N 1.21 S368P 1.05 S368Q 1.10 S368R1.14 S368T 1.15 S368W 1.16 S369A 1.22 S369D 1.05 S369F 1.20 S369G 1.05S369K 1.12 S369L 1.49 S369M 1.36 S369N 1.25 S369P 1.16 S369R 1.12 S369T1.25 N376F 1.12 N376G 1.26 N376H 1.21 N376K 1.40 N376L 1.34 N376P 1.05N376Q 1.11 N376S 1.09 N376V 1.19 N376W 1.12 N376Y 1.05 Y395A 1.05 Y395C1.02 Y395F 1.03 Y395G 1.13 Y395H 1.10 Y395L 1.50 Y395N 1.20 Y395Q 1.18Y395R 1.14 Y395S 1.13 Y395T 1.04 A398C 1.10 A398D 1.39 A398F 1.05 A398G1.17 A398H 1.33 A398I 1.41 A398K 1.47 A398L 1.44 A398N 1.23 A398P 1.38A398Q 1.43 A398R 1.59 A398S 1.14 A398T 1.25 A398V 1.29 A398W 1.45 A398Y1.38 S401A 1.12 S401E 1.08 S401I 1.05 S401N 1.12 S401P 1.15 S401R 1.25S401T 1.26 S401V 1.18 R408A 1.14 R408E 1.41 R408G 1.15 R408H 1.12 R408I1.19 R408K 1.80 R408L 1.55 R408N 1.09 R408Q 1.23 R408S 1.17 N409A 1.25N409C 1.18 N409D 1.21 N409E 1.27 N409F 1.32 N409G 1.14 N409H 1.29 N409I1.56 N409K 1.44 N409L 1.57 N409M 1.17 N409Q 1.03 N409R 1.29 N409V 1.11N409W 1.58 T412L 1.10 S451R 1.01

Example 5 High Performing Variants from an Ethanol Screening Assay

Variants were tested in an ethanol screening assay using the assaysdescribed above. Table 7 shows the results of the screening assay forvariants with a Performance Index (PI)>1.0 compared to the parent TrGAPI. The PI is a measure of specific activity (activity/mg enzyme). ThePI of the specific activity is the quotient “Variant-specificactivity/WT-specific activity.” The PI of the specific activity is 1.0and a variant with a PI>1.0 has a specific activity that is greater thanthe parent TrGA. The specific activity is the activity measured by theethanol screening assay divided by the results obtained in the Bradfordassay described above.

TABLE 7 Ethanol Screening P.I. Variant 32° C., pH 4 D4A 1.07 D4C 1.08D4E 1.23 D4L 1.34 D4R 1.18 D4S 1.17 F5C 1.35 I12L 1.19 I12R 1.13 D24E1.60 D24L 1.19 D24W 1.03 D24Y 1.14 F29A 1.05 F29C 1.12 F29D 1.20 F29E1.05 F29I 1.26 F29L 1.42 F29Q 1.01 F29S 1.07 F29V 1.06 I43D 1.14 I43F1.33 I43R 1.21 I43Y 1.05 D44E 1.37 D44F 1.07 D44G 1.03 D44H 1.11 D44K1.09 D44N 1.07 D44S 1.08 D44Y 1.07 Y70E 1.02 Y70G 1.06 Y70K 1.01 Y70M1.36 Y70P 1.15 Y70R 1.40 Y70S 1.04 Q75A 1.10 Q75K 1.77 R76K 1.06 R76L1.11 R76M 1.13 R76N 1.02 R76T 1.04 R76V 1.05 R76W 1.02 R76Y 1.05 D100A1.08 D100I 1.14 D100L 1.03 D100M 1.12 D100N 1.06 D100P 1.09 D100Q 1.14D100T 1.06 D100W 1.19 D100Y 1.05 N119E 1.02 N119F 1.03 N119Y 1.28 N146C1.11 N146E 1.02 N146G 1.11 N146H 1.07 N146K 1.06 Q148H 1.10 Q148N 1.05Q148V 1.18 Q148W 1.05 Q148Y 1.16 Y169D 1.18 Y169F 1.10 Y169H 1.05 Y169R1.02 Q172E 1.08 Q172G 1.05 Q172R 1.22 Q172S 1.03 F175C 1.18 F175H 1.26F175T 1.28 F175W 1.16 F175Y 1.05 V181F 1.28 V181K 1.35 V181L 1.37 V181R1.01 Q208A 1.22 Q208C 1.17 Q208F 1.12 Q208H 1.02 Q208I 1.02 Q208L 1.32S211A 1.30 S211E 1.30 S211G 1.05 S211L 1.04 S211M 1.05 S211R 1.34 S211W1.07 S211Y 1.08 E243A 1.23 E243L 1.20 E243M 1.26 E243N 1.28 E243R 1.31E243Y 1.25 I292F 1.23 I292H 1.04 I292L 1.21 I292N 1.27 I292R 1.02 I292V1.24 G294A 1.91 G294I 1.92 G294Q 1.99 K297A 1.82 K297D 1.87 K297H 1.79K297M 1.91 K297N 1.87 K297Q 1.85 K297R 1.71 K297S 1.72 K297W 1.70 K297Y1.80 R309L 1.43 Y310F 1.05 Y310Q 1.16 Y310R 1.24 V314D 1.10 V314F 1.04V314H 1.31 V314K 1.08 V314L 1.02 V314N 1.05 V314R 1.06 Y316R 1.42 Y316W1.05 N317H 1.14 N317K 1.02 N317S 1.03 N317T 1.23 K340D 1.33 K340T 1.16K341D 1.04 K341F 1.64 K341G 1.64 K341L 1.04 K341N 1.05 K341S 1.06 T350A1.56 T350D 1.04 T350E 1.59 T350H 1.03 T350N 1.06 T350Q 1.05 T350R 1.02Q356D 1.69 Q356E 1.07 Q356H 1.03 Q356K 1.03 T363A 1.04 T363C 1.54 T363G1.02 T363H 1.09 T363N 1.02 T363R 1.61 T363V 1.05 T363W 1.08 S368D 1.11S368F 1.08 S368H 1.04 S368L 1.07 S368M 1.03 S368N 1.02 S368W 1.24 S369F1.68 S369M 1.04 S369T 1.05 N376G 1.05 N376H 1.10 N376Q 1.16 N376S 1.06N376T 1.12 N376V 1.64 Y395A 1.02 Y395C 1.05 Y395G 1.02 Y395Q 1.63 Y395R1.20 Y395S 1.09 A398D 1.05 A398P 1.03 S401A 1.04 S401D 1.01 S401G 1.04S401N 1.02 S401V 1.06 N409K 1.30 N409L 1.04 N409W 1.31 T412A 1.04 T412G1.06 T412K 1.05 R433Q 1.16 I436A 1.32 I436H 1.02 I436T 1.03 S451A 1.03S451M 1.28 S451T 1.09 S451Y 1.03

Example 6 High Performing Variants from a Sweetener Screening Assay

Variants were tested in a sweetener screening assay as describedhereinabove. Table 8 shows the results of the screening assay whereinvariants with a Performance Index (PI)>1.00 compared to the parent TrGAPI are shown. The PI is a measure of specific activity (activity/mgenzyme). The PI of the specific activity is the quotient“Variant-specific activity/WT-specific activity.” The PI of the specificactivity is 1.0 and a variant with a PI>1.0 has a specific activity thatis greater than the parent TrGA.

TABLE 8 Sweetener screening P.I. 60° C., Variant pH 4.5 D4E 1.09 D4L1.03 D4S 1.07 D24E 1.45 D24L 1.31 D24Y 1.01 I43D 1.05 I43F 1.31 I43R1.28 D44E 1.09 D44H 1.12 D44N 1.31 Y70F 1.26 Y70L 1.22 Q75K 1.12 R76K1.11 R76M 1.03 R76P 1.13 R76T 1.11 R76W 1.07 D100Y 1.04 N119E 1.12 N119Y1.01 N146D 1.05 N146E 1.11 Q148D 1.02 Q148W 1.05 Q172H 1.05 Q172Y 1.03F175H 1.42 F175Y 1.11 V181A 1.10 V181F 1.01 V181K 1.43 V181L 1.42 Q208C1.08 Q208F 1.20 Q208H 1.11 Q208L 1.03 Q208N 1.03 Q208S 1.06 Q208T 1.12S211H 1.16 S211M 1.16 S211R 1.34 S211W 1.09 E243A 1.06 E243F 1.01 E243N1.05 E243R 1.14 E243S 1.09 E243Y 1.07 R245A 1.01 I292N 1.04 I292V 1.12G294A 1.06 G294Q 1.02 K297A 1.04 K297D 1.10 K297Q 1.07 V314D 1.22 V314H1.85 V314K 1.34 V314L 1.13 V314N 1.08 V314R 1.20 V314Y 1.05 Y316R 1.20N317H 1.25 N317K 1.03 K340D 1.21 K340E 1.05 K341D 1.08 K341G 1.22 K341L1.08 K341N 1.08 K341S 1.12 T350H 1.03 T350L 1.04 Q356D 1.31 Q356E 1.04Q356K 1.05 T363C 1.08 T363G 1.04 T363N 1.02 T363R 1.50 S368G 1.04 S368M1.03 N376G 1.02 N376Q 1.07 Y395Q 1.01 A398H 1.03 A398S 1.03 S401A 1.01N409K 1.19 N409T 1.02 N409W 1.01 T412G 1.06 T412S 1.03 I436D 1.02 I436Q1.06 I436T 1.16 S451D 1.01 S451E 1.09 S451F 1.02 S451H 1.11 S451T 1.11

Example 7 Construction of Vectors and Transformation into Trichodermareesei Host Cells A. Construction of Expression Vectors Comprising aPolynucleotide Encoding a Variant GA.

The TrGA expression cassette comprising the DNA sequence SEQ ID NO: 4was cloned into pDONR™201, a Gateway® Entry vector (Invitrogen,Carlsbad, Calif.). The TrGA expression cassette was cloned into theGateway compatible destination vector pTrex3g-DEST (FIG. 5), which isalso described in WO 06/060062, by the Gateway® LR recombinationreaction. The pTrex3g-TrGA expression vector (FIG. 5) enabled theexpression of the TrGA protein (SEQ ID NO: 2) in a Trichoderma reeseihost. Vectors were constructed, which included modified TrGA cDNA codingat least the following variants (1) V314H; (2) S211R; (3) Q208N and (4)Q172F.

B. Transformation.

An expression vector containing a variant GA was transformed into a T.reesei host strain derived from RL-P37 (IA52) and having various genedeletions (Δ cbh1, Δcbh2, Δegl1, Δegl2) using particle bombardment bythe PDS-1000/Helium System (BioRad Cat. No. 165-02257). The protocol isoutlined below, and reference is also made to examples 6 and 11 of WO05/001036.

A suspension of spores (approximately 5×10⁸ spores/ml) from the strainof T. reesei was prepared. One hundred to two hundred microliters ofspore suspension were spread onto the center of plates of Minimal Medium(MM) acetamide medium. The MM acetamide medium had the followingcomposition: 0.6 g/L acetamide; 1.68 g/L CsCl; 20 g/L glucose; 20 g/LKH₂PO₄; 0.6 g/L CaCl₂.2H₂O; 1 ml/L 1000× trace elements solution: 20 g/Lagar; and pH 5.5; 1 ml/L 400× trace element salt solution; citric acid175 g/L, FeSO₄.7H₂O 200 g/L, ZnSO₄.7H₂O 16 g/L, CuSO₄.5H₂O 3.2 g/L,MnSO₄.H₂O 1.4 g/L, H₃BO₃ 0.8 g/L. The spore suspension was allowed todry on the surface of the MM acetamide medium.

Transformation followed the manufacturers instruction. Briefly, 60 mg ofM10 tungsten particles were placed in a microcentrifuge tube. 1 mL ofethanol was added and allowed to stand for 15 minutes. The particleswere centrifuged at 15,000 rpm for 15 seconds. The ethanol was removedand the particles were washed three times with sterile dH₂O before 1 mLof 50% (v/v) sterile glycerol was added. 25 μl of tungsten particlesuspension was placed into a microcentrifuge tube. While continuouslyvortexing, the following were added: 0.5-5 μl (100-200 ng/μl) of plasmidDNA, 25 μl of 2.5 M CaCl₂ and 10 μl of 0.1 M spermidine. The particleswere centrifuged for 3 seconds. The supernatant was removed and theparticles were washed with 200 μl of 70% (v/v) ethanol and centrifugedfor 3 seconds. The supernatant was removed and 24 μl 100% ethanol wasadded, mixed by pipetting, and the tube was placed in an ultrasonicbath, 8 μl aliquots of particles were removed and placed onto the centerof macrocarrier disks that were held in a desiccator. Once thetungsten/DNA suspension had dried the microcarrier disk was placed inthe bombardment chamber along with the plate of MM acetamide with sporesand the bombardment process was performed according to the manufacturersinstructions. After bombardment of the plated spores with thetungsten/DNA particles, the plates were incubated at 28° C. Transformedcolonies were picked to fresh plates of MM acetamide after 4 days(Pentillä et al. (1987) Gene 61: 155-164).

C. Demonstration of GA Activity from the Expressed Variant TrGA inTransformed Cells.

After 5 days growth on MM acetamide plates transformants displayingstable morphology were inoculated into 250 ml shake flasks containing 30ml of Proflo medium. Proflo medium contained: 30 g/L α-lactose; 6.5 g/L(NH₄)₂SO₄; 2 g/L KH₂PO₄; 0.3 g/L MgSO₄.7H₂O; 0.2 g/L CaCl₂.2H₂O; 1 ml/L400× trace element salt solution: citric acid 175 g/L, FeSO₄.7H₂O 200g/L, ZnSO₄.7H₂O 16 g/L, CuSO₄.5H₂O 3.2 g/L, MnSO₄.H₂O 1.4 g/L, H₃BO₃ 0.8g/L; 2 ml/L 10% Tween 80; 22.5 g/L ProFlo cottonseed flour (Tradersprotein, Memphis, Tenn.); 0.72 g/L CaCO₃. After two days growth at 28°C. and 140 rpm, 10% of the Proflo culture was transferred to a 250 mlshake flask containing 30 ml of Lactose Defined Media. The compositionof the Lactose defined Media was as follows 5 g/L (NH₄)₂SO₄; 33 g/L1,4-Piperazinebis(propanesulfonic acid) buffer; 9 g/L casamino acids;4.5 g/L KH₂PO₄; 1.0 g/L MgSO₄.7H₂O; 5 ml/L Mazu DF60-P antifoam (MazurChemicals, IL); 1000× trace element solution; pH 5.5; 40 ml/L of 40%(w/v) lactose solution was added to the medium after sterilization. TheLactose Defined medium shake flasks were incubated at 28° C., 140 rpmfor 4-5 days.

Samples of the culture supernatant were mixed with an appropriate volumeof 2× sample loading buffer with reducing agent. Mycelium was removed bycentrifugation and the supernatant was analyzed for total protein (BCAProtein Assay Kit, Pierce Cat. No. 23225).

GA activity was measured using the p-nitrophenyl-alpha-D-glucopyranoside(pNPG) assay with pNPG as a substrate (Sigma N-1377). In this assay theability of glucoamylase to catalyze the hydrolysis ofp-nitrophenyl-alpha-D-glucopyranoside (pNPG) to glucose andp-nitrophenol is measured. At an alkaline pH, the nitrophenol forms ayellow color that is proportional to glucoamylase activity and ismonitored at 405 nm and compared against an enzyme standard measured asa GAU (Elder, M. T. and Montgomery R. S., Glucoamylase activity inindustrial enzyme preparations using colorimetric enzymatic method,Journal of AOAC International, vol. 78(2), 1995). One GAU is defined asthe amount of enzyme that will produce 1 gm of reducing sugar calculatedas glucose per hour from a soluble starch substrate (4% ds) at pH 4.2and 60° C.

The protein profile was determined by PAGE electrophoresis on NuPAGE®Novex 10% Bis-Tris Gel with MES SDS Running Buffer (Invitrogen,Carlsbad, Calif., USA).

Example 8 Small Scale Applications Testing of Selected Variants onSoluble Starch

Trichoderma reesei host strains expressing the single variants a) V314H,b) S211R, c) Q172F and d) Q208N were grown in fed-batch 14 L fermentorsat 34° C., pH 3.5 in nutrient media including glucose (Cerelose DE99),KH₂PO₄, MgSO₄.7H₂O, (NH₄)₂SO₄, CaCl₂.2H₂O, trace elements and Mazuanti-foam (DF6000K). Upon glucose depletion growth temperature and pHwere shifted to 28° C. and 4.0, respectively. Cell material was removedby filtration and culture supernatants were collected and concentratedto contain greater than 90% glucoamylase as total protein.

Various kinetic properties were determined for glucose production onsoluble potato starch at pH 4.3 at 32° C. and at 60° C. and compared tothe wild-type TrGA. Each of the four variants demonstrated increasedV_(max) (μM glucose/sec) values as compared to the wild type (TrGA)indicating elevated catalytic rates (k_(cat)(sec⁻¹)). FIG. 6 illustratesthe V_(max) of two replicates for each temperature tested.

Example 9 Method to Determine the Performance on EtOH Production ofVariants

Validation of the screening was performed on the variants that wereidentified as having a higher performance index as compared to theparent TrGA (see Table 7/8) using a novel small scale Ethanolapplication test. Twenty-four variants derived from site evaluation andcombinatorial (Table 9) libraries were selected and transformed directlyinto T. reesei for expression and testing on larger scale. The variantswere tested for thermal unfolding using Differential ScanningCalorimetry (DSC analysis described herein below) and performance usinga novel secondary small-scale ethanol application assay. The methodconsisted of two steps: 1) injection of variants onto an anion exchangecolumn to accurately determine the protein concentration; and 2)titration of variants with three different TrGA concentrations(0.3-0.15-0.075 g/28 g ds) in order to calculate their performance onethanol production relative to the wild type molecule.

TABLE 9 List of combinatorial variants variant mutation LR8 Q172F/Q208NLR6 Q172F/Q208N/V314H LR12 Q172F/S211R SW3-1D24E/I43R/D44N/F175H/V181L/V314H/T353R SW3-2D24L/I43F/D44N/F175H/V181L/V314H/T353R ET4-1D24L/Q208Q/I292V/G294A/K297A/Y310R ET4-2D24W/Q208F/I292V/G294Q/K297A/Y310R ET5-1V181L/E243A/I292N/K297N/N317N/Y395Q ET5-2V181L/E243R/I292F/K297A/N317N/Y395Q ET7-1D24Y/V181L/Q208C/G294A/T353R/N375N/N409W ET7-2D24L/V181L/Q208C/G294A/T353R/N375Q/N409W ET8-1D24E/V181K/E243Y/I292V/G294Q/N409K ET8-2D24E/V181F/E243R/I292N/G294I/N409W ET9-1 I043R/E243R/I292F/G294A/K297AET9-2 I043R/E243R/I292L/G294A/K297M

Protein Purification and Determination

A crude enzyme preparation was purified using an AKTA explorer 100 FPLCsystem (Amersham Biosciences, Piscataway, N.J.). β-Cyclodextrin was(Sigma-Aldrich, Zwijndrecht, The Netherlands; 85.608-8) coupled toepoxy-activated Sepharose beads (GE Healthcare, Diegem, Belgium;17-0480-01). The column was used to capture glucoamylases from theenzyme preparation. Enzyme was eluted from the beads using 25 mM Trisbuffer pH 7.5 or 50 mM sodium acetate buffer pH 4.3 containing 10 mMα-cyclodextrin (Sigma, 28705). Purified samples were analyzed bySDS-PAGE. To accurately determine the protein concentration of thevariants an FPLC based protein determination method was developed. Theprotein concentration of the purified marker TrGA molecule was firstdetermined using a standard Bradford protocol (Bio-Rad cat#500-0205).Subsequently, purified samples were injected onto a ResourceQ_(—)1 mlcolumn (GE Healthcare) and enzyme was eluted with 25 mM Tris pH buffercontaining 500 mM NaCl. Peak area was determined and the proteinconcentration was calculated relative to the peak area of the TrGAstandard with known concentration.

Small Scale EtOH Application

Table 10 summarizes the production of ethanol and sugars (DP1, DP2,DP>3) by different combinatorial variants. A sample of corn mashliquefact obtained and diluted to 26% DS using thin stillage. The pH ofthe slurry was adjusted to pH 4.3 using 4N sulphuric acid. A 100 galiquot of mash was placed into a 32° C. water bath and allowed toequilibrate. After 100 μl 400 ppm urea addition, 1 ml purified variantTrGA enzyme sample (150 μg/ml) or purified TrGA (300, 150, 75 μg/ml) wasadded to each corn mash sample. Finally, 333 μl of 30 minutes hydrated15 g in 45 ml DI water solution of Red Star Red yeast (Lesaffre yeastCorp. Milwaukee, Wis.) was added to each sample. Samples were taken at5, 21, 28, 48 and 52 hours and analyzed by HPLC using an Aminex HPX-87Hcolumn 9 (Bio-Rad).

Ethanol and Carbohydrate Determinations

A 2 ml eppendorf centrifuge tube was filled with fermentor beer andcooled on ice for 10 minutes. The sample was centrifuged for 3 minutesat 14.000×g and 500 μl of the supernatant was transferred to a test tubecontaining 50 μl of kill solution (1.1 N sulfuric acid) and allowed tostand for 5 minutes. 5.0 ml of water was added to the test tube and thenfiltered into a 0.22 μm filter plate (multiscreen, Millipore, Amsterdam,the Netherlands) and run on HPLC. Column Temperature: 60° C.; mobilephase: 0.01 N sulfuric acid; flow rate 0.6 ml/min; detector: R1;injection volume: 20 μl. The column separates molecules based on chargeand molecular weight; DP1 (monosaccharides); DP2 (disaccharides); DP3(trisaccharides); DP>3 (oligosaccharides sugars having a degree ofpolymerization greater than 3); succinic acid; lactic acid; glycerol;methanol; ethanol.

DSC Analysis

The melting temperature of purified enzyme samples (0.2-0.4 mg/ml) wasdetermined using Differential Scanning Calorimetry (DSC).

TABLE 10 Production of ethanol and saccharides DP > 3 DP2 DP1 ethanolhours (m/v) % (m/v) % (m/v) % (v/v) % TrGA (0.3 mg) 5.5 3.46 2.70 0.911.02 21.5 3.40 0.50 0.06 6.80 28.5 1.68 1.46 0.07 8.13 46 0.04 0.71 0.0610.21 52.5 0.04 0.45 0.03 10.96 TrGA (0.150 mg) 5.5 3.40 2.43 0.15 1.0021.5 3.78 0.21 0.03 4.23 28.5 3.86 0.20 0.03 5.07 46 2.73 0.52 0.06 7.8652.5 1.70 0.87 0.04 7.92 TrGA (0.075 mg) 5.5 3.43 2.16 −0.01 0.94 21.53.54 0.20 0.03 3.10 28.5 3.43 0.18 0.03 3.14 46 3.93 0.18 0.05 4.65 52.54.01 0.18 0.03 4.79 ET7-1 5.5 3.45 2.53 0.21 1.00 21.5 3.94 0.22 0.044.77 28.5 3.89 0.23 0.04 5.58 46 1.58 1.22 0.06 8.64 52.5 0.62 1.50 0.049.14 LR8 5.5 3.43 2.50 0.17 1.00 21.5 3.96 0.22 0.04 4.79 28.5 3.86 0.210.04 6.21 46 1.27 1.11 0.07 9.17 52.5 0.45 1.24 0.04 8.73 LR12 5.5 3.472.51 0.16 1.05 21.5 3.86 0.22 0.04 4.44 28.5 3.94 0.22 0.04 5.30 46 2.091.08 0.07 8.56 52.5 0.99 1.52 0.04 9.16 LR6 5.5 3.37 2.44 0.18 0.96 21.53.88 0.21 0.04 4.44 28.5 3.90 0.20 0.04 5.10 46 2.44 0.64 0.08 8.59 52.51.27 1.01 0.04 8.97 ET8-1 5.5 3.46 2.53 0.22 0.99 21.5 3.99 0.21 0.044.86 28.5 3.90 0.21 0.04 5.76 46 1.29 1.11 0.08 8.94 52.5 0.47 1.25 0.049.56 ET7-2 5.5 3.57 2.46 0.17 1.02 21.5 4.26 0.21 0.03 4.21 28.5 4.370.20 0.04 5.14 46 3.87 0.27 0.05 7.21 52.5 3.27 0.33 0.03 8.07

Table 11 represents the final ethanol yields and the performance of thevariants at 0.15 mg dosage. The performance was calculated byinterpolation of the 0.3 mg and 0.15 mg values of the TrGA by the valuesof the variants.

TABLE 11 Ethanol Yields EtOH performance relative to variant %(v/v) TrGATrGA 0.3 mg 10.21 TrGA 0.15 mg 7.86 1.00 TrGA 0.075 mg 4.65 ET7-1 8.641.33 LR8 9.17 1.56 LR12 8.56 1.30 LR6 8.59 1.31 ET8-1 8.94 1.46 ET7-27.21 0.72

All combinatorial variants except ET7-2 performed better than TrGA wildtype. LR8 performed the best with a 1.56 improved performance.

Table 12 gives an overview of all single site and combinatorial variantstested using the small-scale ethanol application assay. Variants thatare shaded in Table 12 had a better performance than TrGA and also had ahigher thermal unfolding temperature (dT_(m)).

TABLE 12 Performance and thermal unfolding of variants relative to TrGA

The results showed that Chromatography (FPLC) was a useful tool toaccurately determine the protein concentration. The results also showedthat titration of variants with three TrGA concentrations was a valuablemethod to determine the performance of variants on small scale. Sevenvariants performed better than TrGA wild type (see Table 12) and alsohad a higher thermal unfolding temperature and the variants that did notperform as well as TrGA also had a lower T_(m).

Example 10 Specific Activity Determination of a Selected Set ofCombinatorial and Single Site Variants and Substrate Specificity of LR8

The specific activity of a set of the combinatorial variants and severalsingle site variants that were used to construct combinatorial variantswas analyzed (Table 13). LR8 (PI 1.56 determined with small scaleapplication assay) was further studied with respect to substratespecificity. This was done by setting up an MTP assay to determine theglucose production rates of GA variants and to determine substratespecificity of the LR8 variant. The MTP assay was found to discriminatebetween variants and all variants except ET7-1 showed higher rates thanthe wild-type (wt) Trichoderma reesei glucoamylase. Further, severalvariants (LR8/ET8/Q172F) performed 20-30% better than TrGA. LR8performed better on soluble corn starch and two different samples ofcorn mash liquefact compared to wild-type.

Substrates used in the following experiments were soluble corn starchstock solution prepared as follows: 8 g soluble corn starch (Sigma#S4180) was dissolved in 100 ml milliQ water and heated in a microwavefor 1 minute. The dispersion was boiled for 5 minutes and after coolingthe volume was adjusted to 100 ml. 4% soluble corn starch was preparedby diluting the stock solution 1:1 with 100 mM NaAc buffer pH 4. In oneexperiment, a corn liquefact substrate (NE) was prepared using amoisture analyzer to measure % ds, then substrate was diluted 7.5× with50 mM NaAc to finally obtain 4% ds. The substrate was centrifuged for 5′at 2000×g and the supernatant was filtered with a 0.22 μm filter. Inanother experiment, a corn liquefact substrate (BSE) was prepared in thesame way, except that the substrate was diluted 10× beforecentrifugation.

The enzyme was diluted using the Stock solution of 150 μg enzyme/ml (3μg/180 μl reaction mixture). Solutions were further diluted with 50 mMNaAc pH 4.0 as follows: 300 ng (10×), 200 ng, 150 ng, 100 ng, 75 ng, 50ng, 25 ng, 10 ng/180 PI reaction mixture

The assay was performed as follows: 40 μl 50 mM NaAc pH 4.0, 120 μl 4%soluble corn starch, and 20 μl enzyme were added to each well. Sampleswere incubated for 2 hr at 32° C. 900 rpm and terminated on ice afteraddition of 90 μl 800 mM glycine-NaOH buffer pH 10 for 5 min. The platewas centrifuged for 5 min at 2000 rpm at 15° C. To a fresh plate, 85 μlmilliQ water and 100 μl hexokinase cocktail (II test glucose (HK) kit,Instrumental Laboratory #182507-40) and 20 μl supernatant were added.For a glucose (0-1 mg/ml) calibration line 20 μl glucose stock was addedinstead. Plates were incubated for 10 min at room temperature in thedark followed by absorption measurement at 340 nm using the Spectramax.

TABLE 13 performance relative to wt

The results of the assay to determine the glucose production rates of byGA variants are shown in FIGS. 7 and 8. In these figures, the relativeperformance to TrGA was calculated per amount of enzyme added.Conclusions were drawn from the linear region of the graph at 150 ng ofenzyme. The results in FIGS. 7, 8 and Table 13 showed that LR8, ET8,ET7-2, S211R, Q172F and P94N performed better than wild-type over thelinear range.

To determine the substrate specificity of LR8, the performance of LR8and TrGA wild-type was tested on substrates (soluble corn starch, andthe two corn mash substrates produced in Example 10) used in screeningand application. When analyzed by HPLC, the substrates showed adifference in degree of polymerization (DP) pattern (see FIGS. 9-11). InNE and BSE DP1->=DP4 is present while soluble corn starch consist of atleast four or more glucose molecules. On all substrates LR8 performedbetter than wild-type (see FIGS. 9, 10 and 11).

Example 11 Screening and Characterization of Variants Expressed inTrichoderma reesei Using TrGA Site Evaluation Libraries (SELs) in thepTTT Vector

Additional TrGA variants, particularly variants with substitutionswithin the SBD, were created and screened directly in Trichodermareesei. Similar to Example 1, another ten TrGA site saturatedmutagenesis (SSM) libraries were constructed using the pDONR-TrGA entryvector as a template and primers listed in Table 14. The sites include:N61, G73, L417, T430, A431, E503, Q511, A535, A539, and N563. Amongthese sites, E503, Q511, A535, A539, and N563 are located within thestarch binding domain of the TrGA. Subsequently, the recombination wasperformed with the pTTT-Dest vector (FIG. 14) using the LR CLONASE™ IIenzyme mix according to the protocol supplied by Invitrogen. Therecombination products were transformed into E. coli Max efficiency DH5α(Invitrogen) and plated on 2×TY medium [Bacto Tryptone (Difco) 16 g/L,Bacto Yeast Extract (Difco) 10 g/L, NaCL 5 g/L] supplemented with 100μg/mL ampicillin. After overnight incubation at 37° C., 96 singlecolonies of each library were picked from 2×TY agar plates with 100μg/ml ampicillin and grown for 24 hrs at 37° C. in a MTP containing 200μL 2×TY medium with 100 μg/ml ampicillin. Cultures were used forsequence analyses (ABI3100 sequence analyzer, Applied Biosystems). Eachlibrary contained from 15 to 19 different TrGA variants in the finalexpression vector. These variants were individually transformed into T.reesei.

TABLE 14 Primers used to generate additional TrGA SSM libraries AA-position F/R DNA sequence 5′ to 3′  61 FAGATAGCGCTCTTGTCTTCAAGNNSCTCATCGACCGC  61 R CTTGAAGACAAGAGCGCTATC  73 FCTTCACCGAAACGTACGATGCGNNSCTGCAGCGCCGC  73 R CGCATCGTACGTTTCGGTGAA 417 FCAGCGGCACTCCGCTGTCTGCGNNSCACCTGACGTGGT 417 R CGCAGACAGCGGAGTGCCGCT 430 FGTACGCCTCGTTCTTGACAGCCNNSGCCCGTCGGGCT 430 R GGCTGTCAAGAACGAGGCGTA 431 FCGCCTCGTTCTTGACAGCCACGNNSCGTCGGGCTGGC 431 R CGTGGCTGTCAAGAACGAGGC 503 FCTCCGTGGCCGTCACCTTCCACNNSCTCGTGTCGACACA 503 R GTGGAAGGTGACGGCCACGGA 511F GCTCGTGTCGACACAGTTTGGCNNSACGGTCAAGGTG 511 R GCCAAACTGTGTCGACACGAG 535F GAGCGCCGCCGTGGCTCTGGACNNSGTCAACTATGCCGATA 535 RGTCCAGAGCCACGGCGGCGCTCGTGCT 539 FGGCTCTGGACGCCGTCAACTATNNSGATAACCACCCCCTGT 539 RATAGTTGACGGCGTCCAGAGCCACGGC 563 FCGTCGTGGAGTACAAGTACATCNNSGTGGGCCAAGATGGCTCC 563 RGATGTACTTGTACTCCACGACGTCTCC

The SELs were transformed into T. reesei using the PEG-protoplast method(see, e.g., Pentilla et al. (1987) Gene 61: 155-164). The T. reesei hostis a strain derived from RL-P37 (IA52) and having four gene deletions (Δcbh1, Δcbh2, Δegl1, Δegl2; i.e., “quad-deleted;” see U.S. Pat. No.5,847,276, WO 92/06184, and WO 05/001036). Transformation mixturescontaining up to 600 ng of DNA and 1-5×10⁵ protoplasts in a total volumeof 25 μl were treated with 200 ml of 25% PEG solution, diluted with 2volumes of 1.2 M sorbitol solution, mixed with 3% selective top agaroseMM with acetamide and poured onto 2% selective agarose with acetamideeither in 24 well microtiter plates. The plates were incubated at 28° C.for 5 to 8 days. Spores from the total population of transformantsregenerated on each individual well were harvested from the plates usinga solution of 0.85% NaCl, 0.015% Tween 80. Spore suspensions were usedto inoculate fermentations in 96 well MTPs. In the case of 24 well MTPs,an additional plating step on a fresh 24 well MTP with selectiveacetamide MM was introduced in order to enrich the spore number.

The transformants were fermented in MTPs and the culture supernatantscontaining the expressed protein variants were used for assays. Inbrief, MTPs containing 200 μl of LD-GSM medium were inoculated inquadruplicate with spore suspensions of T. reesei transformantsexpressing TrGA variants (more than 10⁴ spores per well). The plateswere incubated at 28° C. with 230 rpm shaking and 80% humidity for 6days. Culture supernatants were harvested by vacuum filtration. Thesupernatants were used in different assay for screening of variants withimproved properties.

Variants exhibiting a performance index of more than 1.0 for thermalstability, specific activity, and both thermal stability and specificactivity are shown in Tables 15-17.

TABLE 15 Thermal stability screening of additional TrGA variants PI ofthermal Variants stability N061V 1.06 G073F 1.44 G073M 1.01 G073N 1.10G073W 1.36 L417I 1.04 L417K 1.20 L417M 1.05 L417Q 1.04 L417R 1.20 L417V1.07 L417Y 1.01 T430A 1.05 T430E 1.02 T430F 1.06 T430H 1.10 T430I 1.04T430K 1.08 T430M 1.17 T430N 1.13 T430Q 1.05 T430R 1.13 T430S 1.17 T430V1.05 A431I 1.03 A431N 1.03 A431P 1.08 A431R 1.08 A431V 1.03 E503A 1.43E503C 1.39 E503S 1.02 E503T 1.04 E503V 1.68 Q511A 1.11 Q511D 1.12 Q511H1.33 Q511N 1.14 Q511S 1.15 A535D 1.12 A535K 1.24 A535N 1.37 A535P 1.59A535R 1.26 A539E 1.32 A539H 1.17 A539M 1.05 A539R 1.36 A539S 1.30 N563A1.14 N563C 1.49 N563E 1.44 N563I 1.65 N563K 1.77 N563L 1.60 N563Q 1.29N563T 1.31 N563V 1.53

TABLE 16 Specific activity screening of additional TrGA variants. PI ofspecific Variants activity N061D 1.05 N0611 1.21 N061L 1.18 N061Q 1.08N061V 1.11 N061W 1.02 G073C 1.02 G073L 1.07 G073W 1.03 L417A 1.12 L417D1.19 L417E 1.10 L417F 1.08 L417G 1.19 L4171 1.10 L417K 1.02 L417Q 1.04L417R 1.30 L417S 1.05 L417T 1.10 L417V 1.21 L417W 1.05 L417Y 1.10 T430A1.19 T430E 1.15 T430F 1.09 T430G 1.16 T430H 1.15 T4301 1.06 T430K 1.24T430M 1.16 T430N 1.07 T430Q 1.15 T430R 1.04 T430V 1.09 A431C 1.04 A431E1.08 A431H 1.11 A4311 1.20 A431L 1.21 A431M 1.12 A431Q 1.22 A431R 1.11A431S 1.09 A431W 1.04 A431Y 1.13 E503C 1.05 E503D 1.06 E503H 1.01 E503S1.10 E503W 1.04 Q511C 1.07 Q511G 1.06 Q511H 1.05 Q5111 1.10 Q511K 1.09Q511T 1.04 Q511V 1.04 A535E 1.19 A535F 1.06 A535G 1.02 A535K 1.07 A535L1.02 A535N 1.04 A535P 1.14 A535R 1.22 A535S 1.06 A535T 1.04 A535V 1.04A535W 1.09 A535Y 1.13 A539E 1.08 A539M 1.03 A539S 1.02 A539W 1.06 A539R1.22

TABLE 17 Additional TrGA variants displaying both increased thermalstability and specific activity PI of PI of specific thermal Variantsactivity stability N061V 1.11 1.06 G073W 1.03 1.36 L4171 1.10 1.04 L417K1.02 1.20 L417Q 1.04 1.04 L417R 1.30 1.20 L417V 1.21 1.07 L417Y 1.101.01 T430A 1.19 1.05 T430E 1.15 1.02 T430F 1.09 1.06 T430H 1.15 1.10T430I 1.06 1.04 T430K 1.24 1.08 T430M 1.16 1.17 T430N 1.07 1.13 T430Q1.15 1.05 T430R 1.04 1.13 T430V 1.09 1.05 A431I 1.20 1.03 A431R 1.111.08 E503C 1.05 1.39 E503S 1.10 1.02 Q511H 1.05 1.33 A535K 1.07 1.24A535N 1.04 1.37 A535P 1.14 1.59 A535R 1.22 1.26 A539E 1.08 1.32 A539M1.03 1.05 A539R 1.22 1.36 A539S 1.02 1.30

Example 12 Characterization of a Selected Set of Single Site andCombinatorial Variants

Based on the results of Examples 4-6 and 11, a selected set ofcombinatorial variants, and single site variants were furthercharacterized for their alter properties. The selected set includessingle site and combinatorial variants with substitution(s) at: 143,D44, N61, G73, G294, L417, T430, A431, E503, Q511, A535, A539, and/orN563. Variants were purified from large-scale fermentation, and PIs ofthermal stability and specific activities were determined. Specifically,specific activities were determined using different substrates,including DP7, cornstarch, and liquefact. The results are shown in Table18 and 19.

TABLE 18 PIs of a selected set of single site variants, each of which isfrom a 500 ml fermentation P.I. P.I. P.I. P.I. DP7- CornStarch- ThermalLiquefact- Variants FPLC FPLC Stability FPLC N61I 1.16 1.35 1.00 1.66A431L 1.15 1.38 1.18 1.51 L417V 1.18 1.32 1.02 1.40 A431Q 1.06 1.20 0.921.24 G294C 1.01 0.84 0.94 1.23 N563K 1.07 1.12 1.97 1.15 Q511H 1.05 1.091.52 1.13 T430M 1.05 1.15 0.89 1.09 E503A 1.08 1.16 1.40 1.09 I43Q 1.111.24 0.94 1.08 A539R 1.15 1.37 1.43 1.08 I43R 1.03 1.07 1.41 1.07 L417R1.23 1.27 1.51 1.04 T430A 1.13 1.35 1.23 1.04 G73F 1.06 1.06 1.45 1.03D44R 0.97 1.06 1.46 0.98 N563I 1.09 1.22 2.06 0.92 D44C 0.80 0.82 0.960.91 E503V 1.17 1.07 1.66 0.88 A535R 1.09 1.44 1.47 0.85

TABLE 19 PIs of a selected set of combinatorial variants P.I. P.I. CornP.I. P.I. DP7- Starch- Thermal Liquefact- Ferm. Variant SubstitutionsFPLC FPLC Stability FPLC 100 ml C4 G73F/T430A/E503V/Q511H 1.05 1.06 2.571.01 100 ml C9 D44C/G73F/L417R/N563K 0.95 1.00 2.57 1.22 100 ml C11D44C/G73F/N563K 0.83 1.04 2.42 500 ml ALL5 I43R/L417V/E503A/Q511H/A539R1.22 1.48 2.37 1.87 100 ml C15 D44C/L417R/N563K 0.95 1.09 2.30 0.99 100ml C8 D44C/G73F/N563K 0.87 1.05 2.28 500 ml C5 D44C/G73F/N563K 0.90 0.712.16 0.97 500 ml TS1 I43R/T430A/E503V/A535R/N563K 2.13 500 ml C7D44C/G73F/E503V/Q511H 0.90 0.79 2.10 0.89 500 ml ALL1I43Q/D44C/L417V/E503A/Q511H/A539R 1.00 1.27 1.95 1.59 500 ml CS4L417V/T430A/Q511H/A539R/N563I 1.21 1.53 1.95 1.73 100 ml C2I43R/G73F/T430A 1.09 1.25 1.91 1.18 500 ml RB7I43Q/T430A/Q511H/L417V/A539R 1.11 1.36 1.85 2.17 500 ml C1G73F/T430A/Q511H 1.20 1.07 1.84 0.96 500 ml ALL2I43Q/L417V/E503A/Q511H/A539R 1.16 1.52 1.84 1.96 500 ml ALL6I43R/N61I/L417V/E503A/Q511H/A539R 1.06 1.53 1.84 2.24 500 ml ALL8I43R/N61I/L417R/E503A/Q511H/A539R 1.10 1.53 1.84 2.11 500 ml RB20I43Q/Q511H/A539R/T430M/N61I/L417V 1.01 1.55 1.79 1.95 500 ml ALL4I43Q/N61I/L417V/E503A/Q511H/A539R 1.08 1.52 1.79 1.73 500 ml CS3L417V/T430A/Q511H/A535R/N563I 0.98 1.40 1.69 1.78 500 ml RB9I43Q/T430A/Q511H/A431L/E503A 0.93 1.39 1.69 1.55 500 ml RB17I43Q/Q511H/A539R/E503A 1.12 1.38 1.68 1.40 500 ml ALL7I43R/L417R/E503A/A539R 1.04 1.54 1.67 1.62 500 ml TS4I43R/T430A/E503A/Q511H/N563K 0.98 1.33 1.66 1.33 500 ml RB18I43Q/Q511H/A539R/T430M 1.06 1.44 1.64 1.40 100 ml C12 I43R/T430A/ 1.151.35 1.60 1.26 500 ml RB16 I43Q/Q511H/A539R/N61I 1.11 1.43 1.60 1.34 500ml TS5 D44R/T430A/Q511H/A535R 0.89 1.22 1.59 1.21 500 ml RB8I43Q/T430A/Q511H/A431L/A539R 1.02 1.38 1.59 2.18 500 ml RB11I43Q/T430A/Q511H/L417V/A539R/ 1.05 1.36 1.58 2.03 A431L 500 ml CS1L417V/T430A/A431L/Q511H/A535R/ 1.08 1.71 1.56 2.35 A539R/N563I 500 mlLQ5 L417R/A431L/Q511H 0.95 1.21 1.56 1.88 500 ml CS2L417V/T430A/A431Q/Q511H/A535R/ 1.07 1.71 1.56 2.32 A539R/N563I 500 mlTS2 D44R/E503A/Q511H/N563I 0.90 1.14 1.53 1.23 500 ml RB15I43Q/Q511H/A539R 1.19 1.42 1.52 1.69 500 ml RB13 I43Q/Q511H/L417V 1.061.38 1.50 1.50 500 ml RB5 I43Q/T430A/Q511H/A539R 0.99 1.45 1.47 1.59 500ml RB4 I43Q/T430A/Q511H/E503A 1.07 1.29 1.47 1.50 500 ml RB19I43Q/Q511H/A539R/T430M/N61I 1.00 1.50 1.47 1.88 500 ml ALL3I43Q/D44C/N61I/L417V/E503A/ 0.82 1.30 1.43 1.64 Q511H/A539R 500 ml LQ4G294C/L417R/A431Q/Q511H 1.02 1.19 1.43 1.65 500 ml RB2I43Q/T430A/Q511H/L417V 1.09 1.38 1.42 1.87 500 ml TS3 E503A/N563I/ 0.941.17 1.39 1.04 500 ml RB12 I43Q/Q511H/N61I 0.96 1.25 1.37 1.56 500 mlRB14 I43Q/Q511H/A431L 1.01 1.31 1.37 1.55 500 ml LQ6 L417V/A431Q/Q511H0.94 1.33 1.35 2.05 100 ml C14 I43Q/T430A/Q511H 1.09 1.34 1.33 1.46 500ml LQ3 G294C/L417V/A431L/Q511H 0.80 1.21 1.29 2.07 500 ml RB3I43Q/T430A/Q511H/A431L 1.20 1.25 1.29 1.70 500 ml RB10I43Q/T430A/Q511H/N61I/A539R/A431L 0.87 1.47 1.29 1.73 500 ml RB1I43Q/T430A/Q511H/N61I 1.19 1.15 1.27 1.47 100 ml C13 I43Q/T430A/ 1.071.29 1.10 1.45 500 ml LQ1 G294C/L417R/A431L 0.90 1.08 1.04 1.77 500 mlLQ2 G294C/L417V/A431Q 0.80 1.08 1.02 2.08 500 ml RB6I43Q/T430A/Q511H/N61I/A539R 0.94

Example 11 Crystal Structure of TrGA

The complete three dimensional structure of Trichoderma reesei (Hypocreajecorina) glucoamylase (TrGA) was determined at 1.9 Å resolution. Table20 shows the coordinates for the Trichoderma glucoamylase crystalstructure. TrGA was crystallized in an intact form containing 599residues and all post-translational modifications that would normallyoccur in the natural host. The crystal structure was produced andanalyzed as follows:

For protein expression and purification, the gene encoding H. jecorinaGA was cloned and expressed according to the protocols described in theU.S. Pat. No. 7,413,887.

The TrGA protein material used for all crystallization experiments wasinitially purified in one step by anion exchange chromatography asfollows: concentrated culture supernatants of expressed TrGA, consistingof 180 mg/ml total protein, were prepared by diluting sample 1:10 in a25 mM Tris-HCl, pH 8.0 buffer. A HiPrep 16/10 Q Sepharose FF column (GEHealthcare) was employed for the anion exchange purification. The HiPrepcolumn was equilibrated with 4 column volumes (CV) starting buffer (25mM Tris-HCl, pH 8.0) followed by application of 10 ml of the dilutedprotein sample. An 8 CV linear gradient of 0 to 140 mM NaCl in therunning buffer (25 mM Tris-HCl, pH 8.0) was applied to elute boundprotein. Bound TrGA eluted from the HiPrep Q sepharose column at a saltconcentration of approximately 80 mM NaCl. Fractions containing pureTrGA protein were pooled and concentrated to 50 mg/ml using a 25 mlVivaspin centrifugal concentration tube (Viva Science) with a molecularweight cutoff (MWCO) of 10 kD. Purified and concentrated TrGA materialwas buffer exchanged using a DG-10 desalting column (Bio-Rad)equilibrated with 50 mM sodium acetate buffer, pH 4.3. Proteinconcentrations were determined by measuring the absorbance at 280 nm.The initially purified and concentrated TrGA protein stock wasthereafter stored at −20° C.

Two additional purification steps, on additional anion exchangepurification, and a size exclusion purification, were introduced toenhance the crystability of the TrGA protein material. These twoadditional purification steps were performed as follows. In the firstanion exchange purification step a 10 ml MonoQ column (GE Healthcare)was employed. A Sample of 1 ml of the initially purified and frozen TrGAmaterial (50 mg protein) was thawed and the buffer was changed to 20 mMTris-HCl, pH 8.0, by repeated dilution of the sample to 6 ml in the newbuffer, followed by a concentration of the sample again to 0.5 ml usinga 6 ml 5 kD MWCO concentration tube. The TrGA sample was diluted afterthe last concentration step in distilled water until a conductivity ofthe protein sample was reached that corresponded to the conductivity ofthe starting buffer of the anion purification, i.e. 25 mM Tris-HCl, pH8.0. The MonoQ column was first equilibrated with 4 column volumes (CV)starting buffer, followed by application of the diluted protein sampleto the column. Bound protein was eluted from the MonoQ column by twodifferent gradients. In the first a 4 CV linear pH gradient was appliedwhere the pH of the starting buffer was decreased from 8.0 to 6.0. Inthe second gradient an 8 CV long salt gradient was applied in which thesalt concentration was increased from 0 to 350 mM NaCl in the runningbuffer (25 mM Tris-HCl, pH 6.0). Bound TrGA was found to elute from thecolumn during the second salt gradient at an approximate NaClconcentration of 150 mM. Fractions containing TrGA were pooled andconcentrated to 2 ml using a 6 ml 5 kD MWCO Vivaspin concentration tube.The concentrated TrGA sample was thereafter applied to a Superdex 20016/60 size exclusion column (GE Healthcare) equilibrated with 4 CV of 20mM Tris-Cl, pH 8.0, and 50 mM NaCl, which also was used as runningbuffer. Fractions from the main elution peak after the size exclusionpurification were pooled and concentrated to an approximate proteinconcentration of 7.5 mg/ml using a 6 ml 5 kD MWCO Vivaspin concentrationtube.

For protein crystallization, the protein sample that was used to findthe initial TrGA crystallization conditions was a sample of the TrGAmaterial that was purified once by anion exchange purification andthereafter stored at −20° C. The TrGA protein sample was thawed anddiluted with 50 mM sodium acetate buffer, pH 4.3, to approximately 12mg/ml, prior to the initial crystallization experiments. Theorthorhombic X-ray dataset, was used to solve the TrGA structure bymolecular replacement (MR), and the high-resolution orthorhombicdataset, used for the final orthorhombic space group TrGA structuremodel. The orthorhombic TrGA crystals were found to grow in solutionconsisting of 25% PEG 3350, 0.20 M ammonium acetate, 0.10M Bis-Tris pH5.5 (reservoir solution), using the vapor-diffusion method with hangingdrops (McPherson 1982), at 20° C. Crystallization drops were prepared bymixing equal amounts of protein solution (12 mg/ml) and reservoirsolution to a final volume of 10 μl. The TrGA crystals were found tobelong to the orthorhombic space group P212121 with approximate celldimensions: α=52.2 Å, b=99.2 Å, c=121.2 Å, and have a calculated V_(m)of 2.3 (Matthews 1968) with one molecules in the asymmetric unit.

For X-ray data collection, the two orthorhombic TrGA datasets werecollected from single crystals mounted in sealed capillary tubes, atroom temperature. The initial lo-resolution orthorhombic TrGA X-raydataset, used to solve the structure by molecular replacement methods(MR), was collected on a home X-ray source, an MSC/Rigaku (MolecularStructures Corp., The Woodlands, Texas) Raxis IV++ image plate detectorwith focusing mirrors using Cu Kα radiation from a Rigaku RU200 rotatinganode generator. This dataset was processed, scaled, and averaged usingthe d*trek software provided by MSC/Rigaku. The C centered monoclinicdataset was collected from a single frozen TrGA crystal at 100K,equilibrated in a cryo-protective agent comprised of 25% PEG 3350, 15%Glycerol 50 mM CaCl₂ and 0.1 M Bis-Tris pH 5.5 as cryoprotectant,mounted in rayon-fiber loops, and plunge frozen in liquid nitrogen priorto transportation to the synchrotron. The high-resolution orthorhombic(1.9 Å) data set and the C centric monoclinic dataset (1.8 Å) were bothcollected at a synchrotron source, beam line 911:5 at MAX LAB in Lund,Sweden. Both datasets that were collected at a synchrotron source wereprocessed with MOSFLM, and scaled with program SCALA included in theCCP4 program package (Collaborative Computational Project Number 41994). All subsequent data processing was performed using the CCP4program package (Collaborative Computational Project Number 4 1994),unless otherwise stated. A set of 5% of the reflections from each dataset was set aside and used for monitoring the R-free (Brünger, A (1992)Nature, 355:472-475).

The TrGA structure was initially solved by MR with the automaticreplacement program MOLREP (Collaborative Computational Project Number 41994), included in the CCP4 program package, using the initiallo-resolution orthorhombic dataset, and using the coordinates ofAspergillus. awamori GA (AaGA) variant X100 (pdb entry 1GLM (Aleshin etal. (1994) J. Mol. Boil. 238: 575-591) as search model. The A. awamoriGA search model was edited to remove all glycosylation moieties attachedto the protein molecule as N- and O-glycosylations, and all solventmolecules before carrying out the MR experiments. All reflectionsbetween 36.8 and 2.8 Å resolution, from the initial lo resolution TrGAdataset, was used for the MR solution. The MR program found a singlerotation function solution, with a maxima of 11.11 above background, thenext highest maxima was 3.8σ above the background. The translationfunction solution gave an R-factor of 48.7% and had a contrast factor of17.4. The MR solution was refined for 10 cycles of restrained leastsquares refinement using the program Refmac 5.0 (Murshudov et al (1997)Acta Crystallogr. D53: 240-255). This lowered the crystallographicR-factor to 31.1% while the R-free value dropped from 42.2% to 41.1%.

The refined MR solution model was used to calculate an initial densitymap from the lo-resolution orthorhombic TrGA dataset. Electron densityfor a disulfide bridge between residues 19 and 26 of TrGA, a disulfidebridge not present in the A. awamori variant X100 structure model, couldreadily be identified in this electron density map. This was taken as anindication that the electron density map was of sufficient quality to beused to build a structure model of TrGA from its amino acid sequence.The initial TrGA structure model, based on the lo-resolution dataset,was refined with alternating cycles of model building using Coot (Emsleyand Cowtan, (2004) Acta Crystallogr. D boil Crystallogr. 60: 2126-2132),and maximum likelihood refinement using Refmac 5.0.

The resolution of the initial TrGA structure model was extended to theresolution of the high-resolution orthorhombic dataset (1.9 Å) byrefining the initial TrGA structure model against the high-resolutiondataset for 10 cycles of restrained refinement using the program Refmac5.0. Most water molecules in the structure models were locatedautomatically by using the water picking protocols in the refinementprograms, and then manually selected or discarded by inspection by eye.All structural comparisons were made with either Coot (Emsley and Cowtan(2004) supra) or O (Jones et al. (1991) Acta Crystallogr. A47: 110-119),and figures were prepared with PyMOL (Delano W. L. (2002) The PyMOLMolecular Graphics System. Palo Alto, Calif., USA; Delano Scientific).

From these results, it can be seen that the TrGA catalytic core segmentfollowed the same (α/α)₆-barrel topology described by Aleshin et al.1992 for the AaGA, consisting of a double barrel of alpha helices withthe C-terminal of the outer helix leading into the N-terminus of aninner helix. It was possible to identify key differences in the electrondensity such as the disulfide bridge between residues 19 and 26 and aninsertion (residues 257-260) relative to AaGA. The segment comprising80-100 also underwent extensive model rebuilding. One majorglycosylation site was identified at Asn 171, which had up to fourglycoside moieties attached. A similar glycosylation site was identifiedin AaGA. Additionally, the catalytic core containing three cis-peptidesbetween residues 22-23, 44-45 and 122-123 were conserved between TrGAand AaGA. Overall there was an rms variation of 0.535 Å between 409 outof 453 Cα atoms when comparing the coordinates of the catalytic cores ofTrGA and AaGA.

Example 14 Homology Between TrGA and AaGA

The crystal structure of the TrGA identified in Example 13, wassuperposed on the previously identified crystal structure of theAspergillus awamori GA (AaGA). The AaGA crystal structure was obtainedfrom the protein database (PDB) and the form of AaGA that wascrystallized was the form containing only a catalytic domain. Thestructure of the Trichoderma reesei glucoamylase with all three regionsintact was determined to 1.8 Angstrom resolution herein (see Table 15and Example 12). Using the coordinates (see Table 20) the structure wasaligned with the coordinates of the catalytic domain from Aspergillusawamorii strain X100 that was determined previously (Aleshin, A. E.,Hoffman, C., Firsov, L. M., and Honzatko, R. B. 1994 Refined crystalstructures of glucoamylase from Aspergillus awamori var. X100. J MolBiol 238: 575-591 and the PDB). As seen in FIGS. 12 and 13 the structureof the catalytic domain overlapped very closely and allowed theidentification of equivalent residues based on this structuralsuperposition.

Based on this analysis, sites were identified that could be mutated inTrGA and result in increased stability and/or specific activity. Thesesites include 108, 124, 175 and 316 at the active site. Also identifiedwere specific pairwise variants Y47W/Y315F and Y47F/Y315W. Other sitesidentified were 143, D44, P45, D46, R122, R125, V181, E242, Y310, D313,V314, N317, R408, and N409. Because of the high structural homology itis expected that beneficial variants found at sites in TrGA would havesimilar consequences in Aspergillus awamori and other homologousglucoamylases.

Various modifications and variations of the described methods and systemof the disclosure will be apparent to those skilled in the art withoutdeparting from the scope and spirit of the disclosure. Although thedisclosure has been described in connection with specific representativeembodiments, it should be understood that the subject matters as claimedshould not be unduly limited to such specific embodiments. Indeed,various modifications of the described modes for carrying out thedisclosure that are obvious to those skilled in the art are intended tobe within the scope of the following claims.

TABLE 20 CRYST1 52.185 99.232 121.240 90.00 90.00 90.00 ORIGX1 1.0000000.000000 0.000000 0.00000 ORIGX2 0.000000 1.000000 0.000000 0.00000ORIGX3 0.000000 0.000000 1.000000 0.00000 SCALE1 0.019163 −0.000001−0.000001 0.00000 SCALE2 0.000000 0.010077 0.000000 0.00000 SCALE30.000000 0.000000 0.008248 0.00000 ATOM 1 N SER A 1 −30.485 30.567−21.185 1.00 37.11 ATOM 2 CA SER A 1 −30.568 29.350 −20.326 1.00 37.00ATOM 3 CB SER A 1 −31.953 28.707 −20.424 1.00 37.27 ATOM 4 OG SER A 1−32.137 28.089 −21.695 1.00 40.11 ATOM 5 C SER A 1 −29.519 28.345−20.772 1.00 35.91 ATOM 6 O SER A 1 −29.043 28.415 −21.911 1.00 35.46ATOM 7 N VAL A 2 −29.170 27.425 −19.867 1.00 34.51 ATOM 8 CA VAL A 2−28.302 26.293 −20.179 1.00 33.56 ATOM 9 CB VAL A 2 −28.142 25.339−18.955 1.00 33.84 ATOM 10 CG1 VAL A 2 −27.349 24.103 −19.316 1.00 34.20ATOM 11 CG2 VAL A 2 −27.468 26.057 −17.827 1.00 34.79 ATOM 12 C VAL A 2−28.846 25.506 −21.363 1.00 32.48 ATOM 13 O VAL A 2 −28.086 25.109−22.245 1.00 31.10 ATOM 14 N ASP A 3 −30.160 25.286 −21.381 1.00 31.43ATOM 15 CA ASP A 3 −30.791 24.530 −22.457 1.00 31.38 ATOM 16 CB ASP A 3−32.283 24.323 −22.190 1.00 32.17 ATOM 17 CG ASP A 3 −32.522 23.492−20.943 1.00 35.28 ATOM 18 OD1 ASP A 3 −32.413 22.251 −21.028 1.00 36.80ATOM 19 OD2 ASP A 3 −32.786 24.092 −19.870 1.00 40.63 ATOM 20 C ASP A 3−30.556 25.153 −23.818 1.00 30.59 ATOM 21 O ASP A 3 −30.282 24.446−24.778 1.00 30.19 ATOM 22 N ASP A 4 −30.644 26.477 −23.875 1.00 29.89ATOM 23 CA ASP A 4 −30.369 27.244 −25.083 1.00 29.99 ATOM 24 CB ASP A 4−30.601 28.731 −24.822 1.00 31.12 ATOM 25 CG ASP A 4 −32.088 29.121−24.785 1.00 34.16 ATOM 26 OD1 ASP A 4 −32.991 28.260 −24.925 1.00 36.06ATOM 27 OD2 ASP A 4 −32.340 30.332 −24.608 1.00 39.96 ATOM 28 C ASP A 4−28.925 27.049 −25.579 1.00 28.65 ATOM 29 O ASP A 4 −28.697 26.881−26.770 1.00 28.51 ATOM 30 N PHE A 5 −27.961 27.096 −24.660 1.00 26.74ATOM 31 CA PHE A 5 −26.553 26.860 −24.994 1.00 25.21 ATOM 32 CB PHE A 5−25.666 27.110 −23.764 1.00 25.59 ATOM 33 CG PHE A 5 −24.244 26.646−23.931 1.00 26.03 ATOM 34 CD1 PHE A 5 −23.395 27.259 −24.854 1.00 27.29ATOM 35 CE1 PHE A 5 −22.063 26.823 −25.009 1.00 27.33 ATOM 36 CZ PHE A 5−21.593 25.783 −24.228 1.00 26.77 ATOM 37 CE2 PHE A 5 −22.425 25.181−23.286 1.00 28.42 ATOM 38 CD2 PHE A 5 −23.749 25.617 −23.144 1.00 28.42ATOM 39 C PHE A 5 −26.352 25.438 −25.539 1.00 24.23 ATOM 40 O PHE A 5−25.659 25.244 −26.544 1.00 23.56 ATOM 41 N ILE A 6 −26.974 24.458−24.892 1.00 22.71 ATOM 42 CA ILE A 6 −26.835 23.065 −25.312 1.00 22.36ATOM 43 CB ILE A 6 −27.491 22.106 −24.299 1.00 21.86 ATOM 44 CG1 ILE A 6−26.744 22.181 −22.956 1.00 22.27 ATOM 45 CD1 ILE A 6 −27.384 21.347−21.834 1.00 22.36 ATOM 46 CG2 ILE A 6 −27.571 20.669 −24.848 1.00 21.69ATOM 47 C ILE A 6 −27.388 22.855 −26.723 1.00 22.84 ATOM 48 O ILE A 6−26.753 22.216 −27.573 1.00 21.76 ATOM 49 N SER A 7 −28.556 23.420−26.996 1.00 23.10 ATOM 50 CA SER A 7 −29.146 23.175 −28.309 1.00 23.90ATOM 51 CB SER A 7 −30.627 23.570 −28.320 1.00 25.04 ATOM 52 OG SER A 7−30.717 24.982 −28.282 1.00 30.08 ATOM 53 C SER A 7 −28.340 23.874−29.422 1.00 22.78 ATOM 54 O SER A 7 −28.186 23.337 −30.508 1.00 22.94ATOM 55 N THR A 8 −27.800 25.053 −29.140 1.00 22.50 ATOM 56 CA THR A 8−26.984 25.780 −30.115 1.00 23.05 ATOM 57 CB THR A 8 −26.834 27.247−29.698 1.00 23.65 ATOM 58 OG1 THR A 8 −28.138 27.839 −29.700 1.00 27.60ATOM 59 CG2 THR A 8 −25.939 28.018 −30.660 1.00 26.76 ATOM 60 C THR A 8−25.601 25.159 −30.307 1.00 21.46 ATOM 61 O THR A 8 −25.109 25.051−31.437 1.00 21.38 ATOM 62 N GLU A 9 −24.978 24.768 −29.200 1.00 19.11ATOM 63 CA GLU A 9 −23.596 24.269 −29.243 1.00 18.01 ATOM 64 CB GLU A 9−22.959 24.334 −27.847 1.00 17.76 ATOM 65 CG GLU A 9 −21.449 23.945−27.794 1.00 17.71 ATOM 66 CD GLU A 9 −20.536 24.892 −28.609 1.00 20.86ATOM 67 OE1 GLU A 9 −20.949 26.010 −28.971 1.00 19.89 ATOM 68 OE2 GLU A9 −19.389 24.500 −28.909 1.00 19.22 ATOM 69 C GLU A 9 −23.462 22.846−29.784 1.00 17.77 ATOM 70 O GLU A 9 −22.423 22.505 −30.368 1.00 18.05ATOM 71 N THR A 10 −24.485 22.020 −29.593 1.00 15.87 ATOM 72 CA THR A 10−24.404 20.609 −29.958 1.00 17.31 ATOM 73 CB THR A 10 −25.677 19.823−29.525 1.00 17.46 ATOM 74 OG1 THR A 10 −25.768 19.860 −28.090 1.0017.46 ATOM 75 CG2 THR A 10 −25.616 18.374 −30.037 1.00 18.42 ATOM 76 CTHR A 10 −24.026 20.346 −31.430 1.00 17.40 ATOM 77 O THR A 10 −23.07319.615 −31.682 1.00 17.22 ATOM 78 N PRO A 11 −24.764 20.934 −32.412 1.0018.30 ATOM 79 CA PRO A 11 −24.346 20.649 −33.798 1.00 18.11 ATOM 80 CBPRO A 11 −25.440 21.317 −34.662 1.00 18.57 ATOM 81 CG PRO A 11 −26.09422.310 −33.771 1.00 19.16 ATOM 82 CD PRO A 11 −25.975 21.779 −32.3611.00 18.54 ATOM 83 C PRO A 11 −22.963 21.231 −34.142 1.00 17.81 ATOM 84O PRO A 11 −22.241 20.655 −34.964 1.00 17.74 ATOM 85 N ILE A 12 −22.60122.353 −33.520 1.00 16.85 ATOM 86 CA ILE A 12 −21.279 22.936 −33.7311.00 16.66 ATOM 87 CB ILE A 12 −21.161 24.319 −33.112 1.00 17.25 ATOM 88CG1 ILE A 12 −22.194 25.267 −33.751 1.00 19.25 ATOM 89 CD1 ILE A 12−22.289 26.635 −33.101 1.00 21.45 ATOM 90 CG2 ILE A 12 −19.714 24.855−33.270 1.00 18.75 ATOM 91 C ILE A 12 −20.170 22.023 −33.178 1.00 16.30ATOM 92 O ILE A 12 −19.155 21.798 −33.848 1.00 14.64 ATOM 93 N ALA A 13−20.360 21.527 −31.951 1.00 15.28 ATOM 94 CA ALA A 13 −19.375 20.627−31.304 1.00 15.19 ATOM 95 CB ALA A 13 −19.788 20.332 −29.883 1.00 15.31ATOM 96 C ALA A 13 −19.204 19.326 −32.092 1.00 15.37 ATOM 97 O ALA A 13−18.083 18.834 −32.297 1.00 13.56 ATOM 98 N LEU A 14 −20.320 18.743−32.531 1.00 15.13 ATOM 99 CA LEU A 14 −20.225 17.503 −33.285 1.00 16.06ATOM 100 CB LEU A 14 −21.630 16.921 −33.510 1.00 17.33 ATOM 101 CG LEU A14 −21.689 15.563 −34.212 1.00 20.02 ATOM 102 CD1 LEU A 14 −20.94614.460 −33.471 1.00 23.09 ATOM 103 CD2 LEU A 14 −23.150 15.225 −34.3901.00 21.86 ATOM 104 C LEU A 14 −19.506 17.749 −34.623 1.00 15.61 ATOM105 O LEU A 14 −18.651 16.947 −35.039 1.00 14.82 ATOM 106 N ASN A 15−19.853 18.852 −35.285 1.00 15.30 ATOM 107 CA ASN A 15 −19.236 19.228−36.567 1.00 16.34 ATOM 108 CB ASN A 15 −19.848 20.545 −37.073 1.0016.07 ATOM 109 CG ASN A 15 −19.232 21.010 −38.388 1.00 18.31 ATOM 110OD1 ASN A 15 −19.565 20.487 −39.431 1.00 17.60 ATOM 111 ND2 ASN A 15−18.312 21.987 −38.325 1.00 21.40 ATOM 112 C ASN A 15 −17.736 19.450−36.405 1.00 15.35 ATOM 113 O ASN A 15 −16.926 18.954 −37.198 1.00 15.29ATOM 114 N ASN A 16 −17.385 20.180 −35.355 1.00 14.82 ATOM 115 CA ASN A16 −15.992 20.555 −35.144 1.00 15.27 ATOM 116 CB ASN A 16 −15.872 21.693−34.148 1.00 15.41 ATOM 117 CG ASN A 16 −16.276 23.023 −34.737 1.0016.53 ATOM 118 OD1 ASN A 16 −16.517 23.136 −35.954 1.00 18.08 ATOM 119ND2 ASN A 16 −16.326 24.050 −33.896 1.00 16.35 ATOM 120 C ASN A 16−15.159 19.362 −34.723 1.00 14.91 ATOM 121 O ASN A 16 −13.975 19.261−35.099 1.00 15.34 ATOM 122 N LEU A 17 −15.771 18.460 −33.956 1.00 14.29ATOM 123 CA LEU A 17 −15.114 17.191 −33.610 1.00 13.90 ATOM 124 CB LEU A17 −16.003 16.346 −32.672 1.00 13.94 ATOM 125 CG LEU A 17 −15.351 15.065−32.133 1.00 16.81 ATOM 126 CD1 LEU A 17 −15.933 14.708 −30.779 1.0020.06 ATOM 127 CD2 LEU A 17 −15.484 13.880 −33.097 1.00 19.31 ATOM 128 CLEU A 17 −14.763 16.409 −34.880 1.00 14.06 ATOM 129 O LEU A 17 −13.61315.957 −35.073 1.00 12.69 ATOM 130 N LEU A 18 −15.774 16.215 −35.7301.00 13.19 ATOM 131 CA LEU A 18 −15.589 15.441 −36.957 1.00 14.25 ATOM132 CB LEU A 18 −16.952 15.027 −37.545 1.00 13.56 ATOM 133 CG LEU A 18−17.717 14.013 −36.684 1.00 16.49 ATOM 134 CD1 LEU A 18 −19.171 13.874−37.165 1.00 16.33 ATOM 135 CD2 LEU A 18 −17.020 12.647 −36.655 1.0018.51 ATOM 136 C LEU A 18 −14.703 16.132 −38.007 1.00 13.49 ATOM 137 OLEU A 18 −14.083 15.435 −38.820 1.00 14.69 ATOM 138 N CYS A 19 −14.61317.462 −37.964 1.00 13.01 ATOM 139 CA CYS A 19 −13.629 18.223 −38.7601.00 13.22 ATOM 140 CB CYS A 19 −13.796 19.738 −38.556 1.00 14.20 ATOM141 SG CYS A 19 −15.125 20.407 −39.642 1.00 16.22 ATOM 142 C CYS A 19−12.182 17.808 −38.450 1.00 13.86 ATOM 143 O CYS A 19 −11.278 18.043−39.259 1.00 13.42 ATOM 144 N ASN A 20 −11.968 17.219 −37.272 1.00 13.21ATOM 145 CA ASN A 20 −10.594 16.850 −36.830 1.00 13.62 ATOM 146 CB ASN A20 −10.394 17.184 −35.324 1.00 13.52 ATOM 147 CG ASN A 20 −10.242 18.687−35.055 1.00 16.17 ATOM 148 OD1 ASN A 20 −10.035 19.119 −33.897 1.0017.34 ATOM 149 ND2 ASN A 20 −10.343 19.486 −36.090 1.00 11.87 ATOM 150 CASN A 20 −10.262 15.381 −37.116 1.00 13.99 ATOM 151 O ASN A 20 −9.23814.857 −36.646 1.00 14.28 ATOM 152 N VAL A 21 −11.123 14.705 −37.8751.00 13.41 ATOM 153 CA VAL A 21 −10.923 13.287 −38.167 1.00 14.20 ATOM154 CB VAL A 21 −12.177 12.448 −37.827 1.00 14.30 ATOM 155 CG1 VAL A 21−11.953 10.971 −38.189 1.00 15.30 ATOM 156 CG2 VAL A 21 −12.517 12.553−36.312 1.00 14.17 ATOM 157 C VAL A 21 −10.551 13.136 −39.644 1.00 14.35ATOM 158 O VAL A 21 −11.255 13.642 −40.491 1.00 15.68 ATOM 159 N GLY A22 −9.461 12.449 −39.953 1.00 15.67 ATOM 160 CA GLY A 22 −9.061 12.300−41.377 1.00 15.70 ATOM 161 C GLY A 22 −9.843 11.182 −42.060 1.00 17.34ATOM 162 O GLY A 22 −10.453 10.358 −41.397 1.00 17.15 ATOM 163 N PRO A23 −9.806 11.117 −43.404 1.00 18.42 ATOM 164 CA PRO A 23 −9.009 11.946−44.278 1.00 18.20 ATOM 165 CB PRO A 23 −8.716 10.990 −45.446 1.00 18.64ATOM 166 CG PRO A 23 −9.983 10.171 −45.560 1.00 18.81 ATOM 167 CD PRO A23 −10.568 10.092 −44.153 1.00 18.59 ATOM 168 C PRO A 23 −9.761 13.182−44.753 1.00 19.05 ATOM 169 O PRO A 23 −9.183 14.055 −45.426 1.00 19.36ATOM 170 N ASP A 24 −11.034 13.288 −44.385 1.00 18.76 ATOM 171 CA ASP A24 −11.878 14.305 −44.996 1.00 19.39 ATOM 172 CB ASP A 24 −13.015 13.636−45.781 1.00 20.87 ATOM 173 CG ASP A 24 −13.920 12.784 −44.913 1.0024.34 ATOM 174 OD1 ASP A 24 −13.502 12.291 −43.839 1.00 27.70 ATOM 175OD2 ASP A 24 −15.079 12.580 −45.330 1.00 28.83 ATOM 176 C ASP A 24−12.452 15.372 −44.061 1.00 18.08 ATOM 177 O ASP A 24 −13.208 16.245−44.509 1.00 17.78 ATOM 178 N GLY A 25 −12.100 15.331 −42.775 1.00 16.65ATOM 179 CA GLY A 25 −12.634 16.343 −41.852 1.00 16.02 ATOM 180 C GLY A25 −12.152 17.718 −42.292 1.00 15.70 ATOM 181 O GLY A 25 −11.033 17.849−42.811 1.00 16.22 ATOM 182 N CYS A 26 −12.979 18.752 −42.086 1.00 15.10ATOM 183 CA CYS A 26 −12.698 20.078 −42.637 1.00 15.46 ATOM 184 CB CYS A26 −13.899 21.037 −42.475 1.00 15.47 ATOM 185 SG CYS A 26 −14.147 21.739−40.786 1.00 16.91 ATOM 186 C CYS A 26 −11.407 20.731 −42.116 1.00 15.65ATOM 187 O CYS A 26 −10.896 21.643 −42.763 1.00 15.80 ATOM 188 N ARG A27 −10.879 20.259 −40.973 1.00 15.02 ATOM 189 CA ARG A 27 −9.615 20.808−40.443 1.00 14.56 ATOM 190 CB ARG A 27 −9.819 21.480 −39.066 1.00 15.00ATOM 191 CG ARG A 27 −10.695 22.728 −39.164 1.00 15.15 ATOM 192 CD ARG A27 −10.826 23.551 −37.888 1.00 14.30 ATOM 193 NE ARG A 27 −11.874 24.566−38.080 1.00 15.05 ATOM 194 CZ ARG A 27 −13.160 24.420 −37.761 1.0017.96 ATOM 195 NH1 ARG A 27 −13.623 23.293 −37.211 1.00 17.37 ATOM 196NH2 ARG A 27 −14.009 25.415 −38.025 1.00 19.55 ATOM 197 C ARG A 27−8.489 19.776 −40.394 1.00 15.49 ATOM 198 O ARG A 27 −7.389 20.079−39.888 1.00 15.40 ATOM 199 N ALA A 28 −8.768 18.577 −40.910 1.00 15.17ATOM 200 CA ALA A 28 −7.805 17.484 −40.988 1.00 16.06 ATOM 201 CB ALA A28 −8.163 16.374 −39.975 1.00 15.52 ATOM 202 C ALA A 28 −7.744 16.913−42.394 1.00 16.96 ATOM 203 O ALA A 28 −7.453 15.730 −42.581 1.00 17.60ATOM 204 N PHE A 29 −8.028 17.756 −43.380 1.00 17.45 ATOM 205 CA PHE A29 −8.188 17.272 −44.744 1.00 18.68 ATOM 206 CB PHE A 29 −8.728 18.376−45.636 1.00 19.45 ATOM 207 CG PHE A 29 −9.299 17.864 −46.919 1.00 20.86ATOM 208 CD1 PHE A 29 −8.515 17.827 −48.071 1.00 23.76 ATOM 209 CE1 PHEA 29 −9.042 17.343 −49.267 1.00 25.46 ATOM 210 CZ PHE A 29 −10.35716.889 −49.318 1.00 22.85 ATOM 211 CE2 PHE A 29 −11.151 16.924 −48.1801.00 24.78 ATOM 212 CD2 PHE A 29 −10.611 17.408 −46.973 1.00 22.71 ATOM213 C PHE A 29 −6.853 16.783 −45.296 1.00 19.10 ATOM 214 O PHE A 29−5.862 17.501 −45.224 1.00 19.40 ATOM 215 N GLY A 30 −6.830 15.558−45.816 1.00 18.73 ATOM 216 CA GLY A 30 −5.603 15.008 −46.398 1.00 19.00ATOM 217 C GLY A 30 −4.717 14.307 −45.399 1.00 19.69 ATOM 218 O GLY A 30−3.657 13.809 −45.768 1.00 19.61 ATOM 219 N THR A 31 −5.133 14.255−44.123 1.00 18.86 ATOM 220 CA THR A 31 −4.450 13.384 −43.165 1.00 19.14ATOM 221 CB THR A 31 −4.846 13.689 −41.689 1.00 18.79 ATOM 222 OG1 THR A31 −6.265 13.579 −41.559 1.00 18.61 ATOM 223 CG2 THR A 31 −4.410 15.106−41.262 1.00 16.47 ATOM 224 C THR A 31 −4.812 11.925 −43.498 1.00 19.11ATOM 225 O THR A 31 −5.713 11.661 −44.313 1.00 19.69 ATOM 226 N SER A 32−4.107 10.982 −42.881 1.00 19.74 ATOM 227 CA SER A 32 −4.367 9.554−43.094 1.00 20.00 ATOM 228 CB SER A 32 −3.411 8.722 −42.248 1.00 20.73ATOM 229 OG SER A 32 −2.064 8.973 −42.612 1.00 21.56 ATOM 230 C SER A 32−5.806 9.217 −42.704 1.00 20.57 ATOM 231 O SER A 32 −6.334 9.778 −41.7321.00 20.70 ATOM 232 N ALA A 33 −6.443 8.319 −43.452 1.00 19.94 ATOM 233CA ALA A 33 −7.768 7.823 −43.068 1.00 19.61 ATOM 234 CB ALA A 33 −8.2326.729 −44.035 1.00 19.31 ATOM 235 C ALA A 33 −7.764 7.285 −41.637 1.0019.10 ATOM 236 O ALA A 33 −6.906 6.482 −41.264 1.00 19.49 ATOM 237 N GLYA 34 −8.742 7.719 −40.856 1.00 17.74 ATOM 238 CA GLY A 34 −8.878 7.282−39.473 1.00 18.31 ATOM 239 C GLY A 34 −7.988 8.020 −38.473 1.00 18.48ATOM 240 O GLY A 34 −8.050 7.739 −37.271 1.00 19.07 ATOM 241 N ALA A 35−7.173 8.959 −38.937 1.00 17.05 ATOM 242 CA ALA A 35 −6.329 9.723−38.000 1.00 17.17 ATOM 243 CB ALA A 35 −5.167 10.376 −38.730 1.00 17.10ATOM 244 C ALA A 35 −7.173 10.784 −37.271 1.00 16.55 ATOM 245 O ALA A 35−8.174 11.271 −37.808 1.00 17.35 ATOM 246 N VAL A 36 −6.793 11.130−36.051 1.00 15.39 ATOM 247 CA VAL A 36 −7.490 12.198 −35.328 1.00 14.41ATOM 248 CB VAL A 36 −8.142 11.687 −34.031 1.00 15.02 ATOM 249 CG1 VAL A36 −8.903 12.828 −33.349 1.00 16.72 ATOM 250 CG2 VAL A 36 −9.081 10.520−34.336 1.00 16.45 ATOM 251 C VAL A 36 −6.407 13.201 −34.944 1.00 14.36ATOM 252 O VAL A 36 −5.421 12.816 −34.311 1.00 14.44 ATOM 253 N ILE A 37−6.566 14.454 −35.331 1.00 13.68 ATOM 254 CA ILE A 37 −5.614 15.470−34.893 1.00 13.67 ATOM 255 CB ILE A 37 −5.528 16.687 −35.849 1.00 13.66ATOM 256 CG1 ILE A 37 −6.847 17.486 −35.901 1.00 14.31 ATOM 257 CD1 ILEA 37 −6.773 18.712 −36.864 1.00 14.21 ATOM 258 CG2 ILE A 37 −5.15816.214 −37.260 1.00 14.62 ATOM 259 C ILE A 37 −6.041 15.908 −33.505 1.0013.27 ATOM 260 O ILE A 37 −7.235 16.011 −33.224 1.00 12.99 ATOM 261 NALA A 38 −5.081 16.159 −32.630 1.00 13.03 ATOM 262 CA ALA A 38 −5.44516.697 −31.333 1.00 12.81 ATOM 263 CB ALA A 38 −4.235 16.680 −30.3771.00 12.73 ATOM 264 C ALA A 38 −6.046 18.122 −31.497 1.00 12.45 ATOM 265O ALA A 38 −6.939 18.503 −30.775 1.00 12.23 ATOM 266 N SER A 39 −5.55518.870 −32.482 1.00 12.90 ATOM 267 CA SER A 39 −5.973 20.246 −32.7691.00 12.85 ATOM 268 CB SER A 39 −5.512 21.211 −31.657 1.00 12.63 ATOM269 OG SER A 39 −5.312 22.563 −32.108 1.00 12.57 ATOM 270 C SER A 39−5.281 20.593 −34.090 1.00 13.33 ATOM 271 O SER A 39 −4.215 20.043−34.376 1.00 13.48 ATOM 272 N PRO A 40 −5.880 21.500 −34.885 1.00 13.12ATOM 273 CA PRO A 40 −5.248 21.999 −36.108 1.00 13.76 ATOM 274 CB PRO A40 −6.407 22.689 −36.860 1.00 14.41 ATOM 275 CG PRO A 40 −7.386 23.045−35.797 1.00 14.32 ATOM 276 CD PRO A 40 −7.223 22.081 −34.665 1.00 13.18ATOM 277 C PRO A 40 −4.126 23.010 −35.860 1.00 14.27 ATOM 278 O PRO A 40−3.474 23.420 −36.824 1.00 14.43 ATOM 279 N SER A 41 −3.864 23.381−34.599 1.00 13.42 ATOM 280 CA SER A 41 −2.799 24.336 −34.318 1.00 14.56ATOM 281 CB SER A 41 −2.788 24.817 −32.840 1.00 14.40 ATOM 282 OG SER A41 −3.962 25.574 −32.534 1.00 16.91 ATOM 283 C SER A 41 −1.446 23.713−34.676 1.00 14.51 ATOM 284 O SER A 41 −1.123 22.626 −34.218 1.00 13.96ATOM 285 N THR A 42 −0.650 24.433 −35.470 1.00 15.63 ATOM 286 CA THR A42 0.652 23.924 −35.919 1.00 16.17 ATOM 287 CB THR A 42 0.750 23.997−37.458 1.00 17.02 ATOM 288 OG1 THR A 42 0.267 25.283 −37.890 1.00 17.03ATOM 289 CG2 THR A 42 −0.110 22.906 −38.078 1.00 16.03 ATOM 290 C THR A42 1.814 24.732 −35.322 1.00 17.44 ATOM 291 O THR A 42 2.967 24.297−35.382 1.00 17.10 ATOM 292 N ILE A 43 1.509 25.913 −34.787 1.00 18.37ATOM 293 CA ILE A 43 2.510 26.786 −34.171 1.00 20.62 ATOM 294 CB ILE A43 2.923 27.952 −35.114 1.00 20.73 ATOM 295 CG1 ILE A 43 3.550 27.428−36.411 1.00 21.88 ATOM 296 CD1 ILE A 43 3.788 28.507 −37.508 1.00 22.99ATOM 297 CG2 ILE A 43 3.895 28.910 −34.409 1.00 21.41 ATOM 298 C ILE A43 1.908 27.395 −32.935 1.00 21.00 ATOM 299 O ILE A 43 0.796 27.921−32.995 1.00 21.76 ATOM 300 N ASP A 44 2.683 27.470 −31.874 1.00 21.61ATOM 301 CA ASP A 44 2.237 27.975 −30.572 1.00 23.04 ATOM 302 CB ASP A44 2.408 29.506 −30.492 1.00 24.75 ATOM 303 CG ASP A 44 2.170 30.064−29.098 1.00 31.28 ATOM 304 OD1 ASP A 44 2.362 29.340 −28.094 1.00 37.92ATOM 305 OD2 ASP A 44 1.766 31.260 −28.997 1.00 40.00 ATOM 306 C ASP A44 0.782 27.608 −30.196 1.00 21.65 ATOM 307 O ASP A 44 −0.046 28.502−29.981 1.00 22.69 ATOM 308 N PRO A 45 0.441 26.449 −29.805 1.00 19.86ATOM 309 CA PRO A 45 1.356 25.320 −29.775 1.00 18.66 ATOM 310 CB PRO A45 0.883 24.549 −28.549 1.00 18.40 ATOM 311 CG PRO A 45 −0.653 24.763−28.586 1.00 18.13 ATOM 312 CD PRO A 45 −0.899 26.066 −29.318 1.00 20.04ATOM 313 C PRO A 45 1.253 24.454 −31.026 1.00 17.74 ATOM 314 O PRO A 450.368 24.652 −31.858 1.00 17.36 ATOM 315 N ASP A 46 2.178 23.512 −31.1601.00 15.95 ATOM 316 CA ASP A 46 2.124 22.573 −32.275 1.00 14.75 ATOM 317CB ASP A 46 3.551 22.255 −32.738 1.00 14.59 ATOM 318 CG ASP A 46 3.60121.161 −33.818 1.00 16.17 ATOM 319 OD1 ASP A 46 2.543 20.787 −34.3891.00 15.61 ATOM 320 OD2 ASP A 46 4.712 20.641 −34.054 1.00 20.18 ATOM321 C ASP A 46 1.436 21.303 −31.748 1.00 13.83 ATOM 322 O ASP A 46 2.08120.489 −31.089 1.00 13.59 ATOM 323 N TYR A 47 0.126 21.165 −31.992 1.0011.86 ATOM 324 CA TYR A 47 −0.621 19.975 −31.580 1.00 12.11 ATOM 325 CBTYR A 47 −1.895 20.387 −30.854 1.00 11.69 ATOM 326 CG TYR A 47 −1.77320.722 −29.377 1.00 12.59 ATOM 327 CD1 TYR A 47 −0.589 21.236 −28.8271.00 13.54 ATOM 328 CE1 TYR A 47 −0.524 21.586 −27.462 1.00 12.81 ATOM329 CZ TYR A 47 −1.652 21.407 −26.673 1.00 13.40 ATOM 330 OH TYR A 47−1.620 21.755 −25.354 1.00 13.08 ATOM 331 CE2 TYR A 47 −2.825 20.887−27.208 1.00 12.02 ATOM 332 CD2 TYR A 47 −2.876 20.540 −28.532 1.0012.76 ATOM 333 C TYR A 47 −0.994 19.090 −32.772 1.00 11.46 ATOM 334 OTYR A 47 −1.885 18.239 −32.692 1.00 11.66 ATOM 335 N TYR A 48 −0.31619.301 −33.893 1.00 12.26 ATOM 336 CA TYR A 48 −0.697 18.639 −35.1321.00 12.80 ATOM 337 CB TYR A 48 −0.323 19.509 −36.348 1.00 12.75 ATOM338 CG TYR A 48 −1.134 19.146 −37.569 1.00 13.21 ATOM 339 CD1 TYR A 48−2.492 19.482 −37.652 1.00 14.46 ATOM 340 CE1 TYR A 48 −3.254 19.124−38.767 1.00 15.86 ATOM 341 CZ TYR A 48 −2.643 18.453 −39.823 1.00 14.62ATOM 342 OH TYR A 48 −3.390 18.106 −40.936 1.00 16.20 ATOM 343 CE2 TYR A48 −1.295 18.086 −39.756 1.00 15.86 ATOM 344 CD2 TYR A 48 −0.543 18.456−38.638 1.00 13.44 ATOM 345 C TYR A 48 −0.072 17.245 −35.187 1.00 13.47ATOM 346 O TYR A 48 0.877 16.986 −35.940 1.00 13.95 ATOM 347 N TYR A 49−0.592 16.360 −34.338 1.00 13.13 ATOM 348 CA TYR A 49 −0.131 14.987−34.171 1.00 13.51 ATOM 349 CB TYR A 49 0.887 14.842 −33.009 1.00 13.11ATOM 350 CG TYR A 49 2.133 15.662 −33.216 1.00 13.90 ATOM 351 CD1 TYR A49 3.193 15.174 −33.996 1.00 13.54 ATOM 352 CE1 TYR A 49 4.354 15.964−34.216 1.00 13.41 ATOM 353 CZ TYR A 49 4.419 17.225 −33.665 1.00 14.69ATOM 354 OH TYR A 49 5.511 18.016 −33.883 1.00 17.21 ATOM 355 CE2 TYR A49 3.365 17.737 −32.906 1.00 13.49 ATOM 356 CD2 TYR A 49 2.227 16.952−32.698 1.00 13.78 ATOM 357 C TYR A 49 −1.349 14.181 −33.783 1.00 13.93ATOM 358 O TYR A 49 −2.390 14.759 −33.406 1.00 13.00 ATOM 359 N MET A 50−1.203 12.857 −33.839 1.00 13.66 ATOM 360 CA MET A 50 −2.241 11.940−33.365 1.00 14.56 ATOM 361 CB MET A 50 −2.447 10.822 −34.381 1.00 15.21ATOM 362 CG MET A 50 −3.532 9.811 −33.947 1.00 15.64 ATOM 363 SD MET A50 −3.996 8.804 −35.361 1.00 19.52 ATOM 364 CE MET A 50 −5.204 7.742−34.566 1.00 17.12 ATOM 365 C MET A 50 −1.797 11.323 −32.060 1.00 14.38ATOM 366 O MET A 50 −0.806 10.583 −32.024 1.00 13.80 ATOM 367 N TRP A 51−2.528 11.608 −30.984 1.00 13.47 ATOM 368 CA TRP A 51 −2.265 10.965−29.720 1.00 13.13 ATOM 369 CB TRP A 51 −2.598 11.930 −28.585 1.00 12.85ATOM 370 CG TRP A 51 −1.478 12.809 −28.116 1.00 13.64 ATOM 371 CD1 TRP A51 −0.671 12.604 −27.011 1.00 13.49 ATOM 372 NE1 TRP A 51 0.211 13.657−26.864 1.00 12.36 ATOM 373 CE2 TRP A 51 −0.023 14.573 −27.858 1.0011.83 ATOM 374 CD2 TRP A 51 −1.076 14.065 −28.674 1.00 13.12 ATOM 375CE3 TRP A 51 −1.506 14.825 −29.772 1.00 11.07 ATOM 376 CZ3 TRP A 51−0.859 16.061 −30.035 1.00 12.87 ATOM 377 CH2 TRP A 51 0.193 16.522−29.218 1.00 13.24 ATOM 378 CZ2 TRP A 51 0.618 15.806 −28.127 1.00 12.61ATOM 379 C TRP A 51 −3.136 9.732 −29.575 1.00 13.35 ATOM 380 O TRP A 51−4.322 9.769 −29.907 1.00 12.89 ATOM 381 N THR A 52 −2.576 8.652 −29.0241.00 13.20 ATOM 382 CA THR A 52 −3.386 7.462 −28.753 1.00 13.02 ATOM 383CB THR A 52 −2.520 6.300 −28.235 1.00 13.66 ATOM 384 OG1 THR A 52 −1.5535.999 −29.246 1.00 15.07 ATOM 385 CG2 THR A 52 −3.341 5.026 −27.952 1.0012.21 ATOM 386 C THR A 52 −4.533 7.807 −27.800 1.00 12.48 ATOM 387 O THRA 52 −5.670 7.402 −28.034 1.00 12.78 ATOM 388 N ARG A 53 −4.224 8.556−26.747 1.00 12.03 ATOM 389 CA ARG A 53 −5.238 8.868 −25.737 1.00 11.89ATOM 390 CB ARG A 53 −4.607 9.570 −24.545 1.00 11.46 ATOM 391 CG ARG A53 −5.611 10.330 −23.618 1.00 13.19 ATOM 392 CD ARG A 53 −4.896 10.881−22.375 1.00 11.14 ATOM 393 NE ARG A 53 −3.793 11.694 −22.819 1.00 12.52ATOM 394 CZ ARG A 53 −2.509 11.330 −22.769 1.00 13.67 ATOM 395 NH1 ARG A53 −2.148 10.182 −22.180 1.00 13.97 ATOM 396 NH2 ARG A 53 −1.590 12.151−23.270 1.00 13.05 ATOM 397 C ARG A 53 −6.395 9.709 −26.319 1.00 12.45ATOM 398 O ARG A 53 −7.558 9.289 −26.244 1.00 11.74 ATOM 399 N ASP A 54−6.090 10.885 −26.874 1.00 11.73 ATOM 400 CA ASP A 54 −7.169 11.747−27.385 1.00 11.90 ATOM 401 CB ASP A 54 −6.638 13.018 −28.053 1.00 12.25ATOM 402 CG ASP A 54 −5.794 13.879 −27.120 1.00 13.97 ATOM 403 OD1 ASP A54 −4.983 13.332 −26.354 1.00 13.57 ATOM 404 OD2 ASP A 54 −5.910 15.110−27.215 1.00 13.88 ATOM 405 C ASP A 54 −8.002 11.005 −28.420 1.00 12.00ATOM 406 O ASP A 54 −9.236 11.138 −28.454 1.00 10.97 ATOM 407 N SER A 55−7.334 10.250 −29.297 1.00 11.19 ATOM 408 CA SER A 55 −8.034 9.544−30.388 1.00 12.36 ATOM 409 CB SER A 55 −7.017 8.901 −31.340 1.00 13.05ATOM 410 OG SER A 55 −6.171 9.930 −31.882 1.00 14.23 ATOM 411 C SER A 55−8.996 8.489 −29.838 1.00 12.57 ATOM 412 O SER A 55 −10.130 8.348−30.327 1.00 12.76 ATOM 413 N ALA A 56 −8.556 7.764 −28.819 1.00 12.60ATOM 414 CA ALA A 56 −9.373 6.718 −28.218 1.00 13.25 ATOM 415 CB ALA A56 −8.517 5.830 −27.329 1.00 12.73 ATOM 416 C ALA A 56 −10.551 7.301−27.415 1.00 13.85 ATOM 417 O ALA A 56 −11.640 6.723 −27.409 1.00 14.51ATOM 418 N LEU A 57 −10.328 8.420 −26.723 1.00 14.23 ATOM 419 CA LEU A57 −11.417 9.059 −25.954 1.00 13.95 ATOM 420 CB LEU A 57 −10.891 10.186−25.060 1.00 14.45 ATOM 421 CG LEU A 57 −10.088 9.751 −23.834 1.00 14.89ATOM 422 CD1 LEU A 57 −9.507 11.013 −23.161 1.00 16.19 ATOM 423 CD2 LEUA 57 −10.919 8.911 −22.867 1.00 16.02 ATOM 424 C LEU A 57 −12.483 9.609−26.886 1.00 13.92 ATOM 425 O LEU A 57 −13.685 9.488 −26.627 1.00 13.70ATOM 426 N VAL A 58 −12.027 10.199 −27.975 1.00 13.15 ATOM 427 CA VAL A58 −12.920 10.751 −28.989 1.00 15.20 ATOM 428 CB VAL A 58 −12.133 11.605−30.031 1.00 14.52 ATOM 429 CG1 VAL A 58 −12.970 11.861 −31.302 1.0017.86 ATOM 430 CG2 VAL A 58 −11.704 12.954 −29.393 1.00 15.79 ATOM 431 CVAL A 58 −13.704 9.624 −29.655 1.00 15.20 ATOM 432 O VAL A 58 −14.9309.718 −29.784 1.00 15.30 ATOM 433 N PHE A 59 −13.026 8.553 −30.058 1.0015.02 ATOM 434 CA PHE A 59 −13.766 7.477 −30.713 1.00 15.23 ATOM 435 CBPHE A 59 −12.882 6.601 −31.582 1.00 15.78 ATOM 436 CG PHE A 59 −12.8597.058 −33.003 1.00 15.10 ATOM 437 CD1 PHE A 59 −11.872 7.927 −33.4441.00 16.45 ATOM 438 CE1 PHE A 59 −11.861 8.401 −34.768 1.00 19.37 ATOM439 CZ PHE A 59 −12.876 8.026 −35.644 1.00 16.44 ATOM 440 CE2 PHE A 59−13.901 7.165 −35.186 1.00 16.90 ATOM 441 CD2 PHE A 59 −13.895 6.709−33.882 1.00 16.17 ATOM 442 C PHE A 59 −14.674 6.681 −29.785 1.00 15.69ATOM 443 O PHE A 59 −15.699 6.175 −30.220 1.00 15.46 ATOM 444 N LYS A 60−14.321 6.586 −28.510 1.00 15.65 ATOM 445 CA LYS A 60 −15.257 5.994−27.552 1.00 16.61 ATOM 446 CB LYS A 60 −14.661 5.954 −26.144 1.00 16.44ATOM 447 CG LYS A 60 −15.626 5.363 −25.059 1.00 17.65 ATOM 448 CD LYS A60 −16.174 3.992 −25.433 1.00 18.35 ATOM 449 CE LYS A 60 −16.738 3.234−24.199 1.00 19.79 ATOM 450 NZ LYS A 60 −17.819 3.976 −23.512 1.00 18.40ATOM 451 C LYS A 60 −16.577 6.779 −27.579 1.00 16.72 ATOM 452 O LYS A 60−17.663 6.182 −27.681 1.00 17.08 ATOM 453 N ASN A 61 −16.487 8.101−27.508 1.00 16.77 ATOM 454 CA ASN A 61 −17.680 8.948 −27.628 1.00 18.06ATOM 455 CB ASN A 61 −17.278 10.424 −27.573 1.00 19.41 ATOM 456 CG ASN A61 −18.465 11.375 −27.643 1.00 22.93 ATOM 457 OD1 ASN A 61 −19.58511.057 −27.231 1.00 30.05 ATOM 458 ND2 ASN A 61 −18.206 12.568 −28.1301.00 29.54 ATOM 459 C ASN A 61 −18.480 8.659 −28.907 1.00 17.28 ATOM 460O ASN A 61 −19.697 8.475 −28.852 1.00 18.11 ATOM 461 N LEU A 62 −17.7998.647 −30.056 1.00 16.54 ATOM 462 CA LEU A 62 −18.460 8.379 −31.334 1.0016.19 ATOM 463 CB LEU A 62 −17.479 8.572 −32.499 1.00 16.85 ATOM 464 CGLEU A 62 −17.047 10.027 −32.697 1.00 18.47 ATOM 465 CD1 LEU A 62 −16.11810.153 −33.916 1.00 20.38 ATOM 466 CD2 LEU A 62 −18.263 10.925 −32.8371.00 19.93 ATOM 467 C LEU A 62 −19.089 6.991 −31.371 1.00 16.01 ATOM 468O LEU A 62 −20.225 6.833 −31.842 1.00 15.98 ATOM 469 N ILE A 63 −18.3875.998 −30.831 1.00 15.67 ATOM 470 CA ILE A 63 −18.910 4.628 −30.810 1.0015.86 ATOM 471 CB ILE A 63 −17.803 3.610 −30.372 1.00 15.88 ATOM 472 CG1ILE A 63 −16.756 3.466 −31.478 1.00 14.98 ATOM 473 CD1 ILE A 63 −15.3752.976 −30.966 1.00 15.62 ATOM 474 CG2 ILE A 63 −18.398 2.251 −30.0161.00 15.96 ATOM 475 C ILE A 63 −20.156 4.538 −29.920 1.00 16.39 ATOM 476O ILE A 63 −21.137 3.854 −30.272 1.00 16.90 ATOM 477 N ASP A 64 −20.1295.242 −28.796 1.00 16.51 ATOM 478 CA ASP A 64 −21.299 5.324 −27.922 1.0017.76 ATOM 479 CB ASP A 64 −20.953 6.022 −26.594 1.00 17.81 ATOM 480 CGASP A 64 −20.089 5.164 −25.682 1.00 17.84 ATOM 481 OD1 ASP A 64 −19.8833.944 −25.963 1.00 18.57 ATOM 482 OD2 ASP A 64 −19.595 5.699 −24.6591.00 20.95 ATOM 483 C ASP A 64 −22.492 5.982 −28.617 1.00 18.64 ATOM 484O ASP A 64 −23.617 5.493 −28.507 1.00 20.85 ATOM 485 N ARG A 65 −22.2627.070 −29.348 1.00 19.57 ATOM 486 CA ARG A 65 −23.341 7.750 −30.097 1.0020.59 ATOM 487 CB ARG A 65 −22.823 9.046 −30.733 1.00 20.62 ATOM 488 CGARG A 65 −22.465 10.083 −29.693 1.00 25.00 ATOM 489 CD ARG A 65 −22.01011.385 −30.324 1.00 28.44 ATOM 490 NE ARG A 65 −23.106 12.071 −30.9901.00 31.14 ATOM 491 CZ ARG A 65 −23.968 12.878 −30.373 1.00 32.75 ATOM492 NH1 ARG A 65 −23.873 13.095 −29.060 1.00 32.10 ATOM 493 NH2 ARG A 65−24.928 13.459 −31.080 1.00 32.31 ATOM 494 C ARG A 65 −23.907 6.841−31.184 1.00 20.83 ATOM 495 O ARG A 65 −25.129 6.711 −31.357 1.00 20.48ATOM 496 N PHE A 66 −22.998 6.213 −31.910 1.00 20.52 ATOM 497 CA PHE A66 −23.340 5.271 −32.966 1.00 21.42 ATOM 498 CB PHE A 66 −22.046 4.778−33.604 1.00 21.97 ATOM 499 CG PHE A 66 −22.224 3.603 −34.520 1.00 21.97ATOM 500 CD1 PHE A 66 −22.601 3.791 −35.844 1.00 23.23 ATOM 501 CE1 PHEA 66 −22.768 2.690 −36.699 1.00 22.87 ATOM 502 CZ PHE A 66 −22.552 1.409−36.221 1.00 22.60 ATOM 503 CE2 PHE A 66 −22.165 1.216 −34.895 1.0023.74 ATOM 504 CD2 PHE A 66 −22.006 2.309 −34.054 1.00 23.07 ATOM 505 CPHE A 66 −24.152 4.084 −32.415 1.00 21.98 ATOM 506 O PHE A 66 −25.0403.551 −33.099 1.00 21.80 ATOM 507 N THR A 67 −23.831 3.654 −31.195 1.0022.48 ATOM 508 CA THR A 67 −24.546 2.537 −30.576 1.00 23.79 ATOM 509 CBTHR A 67 −23.809 1.999 −29.333 1.00 23.68 ATOM 510 OG1 THR A 67 −22.5511.439 −29.745 1.00 23.93 ATOM 511 CG2 THR A 67 −24.613 0.881 −28.6531.00 23.90 ATOM 512 C THR A 67 −25.992 2.925 −30.258 1.00 24.77 ATOM 513O THR A 67 −26.893 2.090 −30.349 1.00 25.31 ATOM 514 N GLU A 68 −26.2074.189 −29.916 1.00 25.62 ATOM 515 CA GLU A 68 −27.540 4.688 −29.616 1.0027.41 ATOM 516 CB GLU A 68 −27.466 6.038 −28.894 1.00 28.13 ATOM 517 CGGLU A 68 −26.997 5.951 −27.446 1.00 32.86 ATOM 518 CD GLU A 68 −28.0955.487 −26.468 1.00 38.33 ATOM 519 OE1 GLU A 68 −29.241 5.982 −26.5421.00 40.42 ATOM 520 OE2 GLU A 68 −27.799 4.633 −25.607 1.00 42.48 ATOM521 C GLU A 68 −28.418 4.784 −30.873 1.00 27.86 ATOM 522 O GLU A 68−29.602 4.429 −30.845 1.00 28.00 ATOM 523 N THR A 69 −27.833 5.260−31.968 1.00 27.32 ATOM 524 CA THR A 69 −28.540 5.373 −33.241 1.00 27.32ATOM 525 CB THR A 69 −29.113 6.792 −33.451 1.00 27.49 ATOM 526 OG1 THR A69 −29.922 7.153 −32.334 1.00 30.86 ATOM 527 CG2 THR A 69 −29.945 6.843−34.719 1.00 29.06 ATOM 528 C THR A 69 −27.563 5.133 −34.359 1.00 26.07ATOM 529 O THR A 69 −26.619 5.905 −34.523 1.00 25.25 ATOM 530 N TYR A 70−27.790 4.064 −35.123 1.00 25.65 ATOM 531 CA TYR A 70 −26.948 3.738−36.267 1.00 25.36 ATOM 532 CB TYR A 70 −27.480 2.504 −37.013 1.00 25.26ATOM 533 CG TYR A 70 −26.638 2.104 −38.217 1.00 25.62 ATOM 534 CD1 TYR A70 −26.949 2.567 −39.498 1.00 25.62 ATOM 535 CE1 TYR A 70 −26.190 2.201−40.611 1.00 26.48 ATOM 536 CZ TYR A 70 −25.099 1.360 −40.437 1.00 25.36ATOM 537 OH TYR A 70 −24.354 0.995 −41.520 1.00 24.80 ATOM 538 CE2 TYR A70 −24.770 0.883 −39.175 1.00 25.59 ATOM 539 CD2 TYR A 70 −25.538 1.259−38.071 1.00 25.36 ATOM 540 C TYR A 70 −26.816 4.909 −37.230 1.00 25.38ATOM 541 O TYR A 70 −27.802 5.573 −37.583 1.00 24.87 ATOM 542 N ASP A 71−25.575 5.127 −37.666 1.00 25.16 ATOM 543 CA ASP A 71 −25.188 6.210−38.550 1.00 25.40 ATOM 544 CB ASP A 71 −24.668 7.404 −37.724 1.00 25.60ATOM 545 CG ASP A 71 −24.361 8.642 −38.573 1.00 26.17 ATOM 546 OD1 ASP A71 −23.801 8.526 −39.681 1.00 25.82 ATOM 547 OD2 ASP A 71 −24.675 9.755−38.108 1.00 27.85 ATOM 548 C ASP A 71 −24.061 5.619 −39.386 1.00 25.64ATOM 549 O ASP A 71 −22.956 5.377 −38.875 1.00 24.82 ATOM 550 N ALA A 72−24.347 5.379 −40.665 1.00 24.95 ATOM 551 CA ALA A 72 −23.380 4.764−41.586 1.00 24.24 ATOM 552 CB ALA A 72 −24.047 4.434 −42.921 1.00 24.35ATOM 553 C ALA A 72 −22.152 5.657 −41.812 1.00 24.04 ATOM 554 O ALA A 72−21.054 5.159 −42.086 1.00 23.21 ATOM 555 N GLY A 73 −22.356 6.970−41.695 1.00 23.63 ATOM 556 CA GLY A 73 −21.265 7.938 −41.761 1.00 24.20ATOM 557 C GLY A 73 −20.285 7.809 −40.596 1.00 23.85 ATOM 558 O GLY A 73−19.067 7.927 −40.806 1.00 24.81 ATOM 559 N LEU A 74 −20.798 7.588−39.376 1.00 22.78 ATOM 560 CA LEU A 74 −19.927 7.347 −38.232 1.00 22.21ATOM 561 CB LEU A 74 −20.662 7.449 −36.879 1.00 22.58 ATOM 562 CG LEU A74 −21.132 8.846 −36.434 1.00 24.06 ATOM 563 CD1 LEU A 74 −21.732 8.793−35.019 1.00 22.31 ATOM 564 CD2 LEU A 74 −20.002 9.869 −36.503 1.0026.04 ATOM 565 C LEU A 74 −19.256 5.999 −38.370 1.00 21.94 ATOM 566 OLEU A 74 −18.060 5.872 −38.098 1.00 20.62 ATOM 567 N GLN A 75 −20.0194.988 −38.814 1.00 21.47 ATOM 568 CA GLN A 75 −19.451 3.654 −38.989 1.0021.07 ATOM 569 CB GLN A 75 −20.469 2.709 −39.619 1.00 21.44 ATOM 570 CGGLN A 75 −20.002 1.280 −39.594 1.00 23.11 ATOM 571 CD GLN A 75 −21.1010.312 −39.945 1.00 24.74 ATOM 572 OE1 GLN A 75 −21.273 −0.719 −39.2901.00 26.57 ATOM 573 NE2 GLN A 75 −21.872 0.654 −40.950 1.00 23.71 ATOM574 C GLN A 75 −18.219 3.704 −39.889 1.00 21.05 ATOM 575 O GLN A 75−17.229 3.046 −39.611 1.00 21.10 ATOM 576 N ARG A 76 −18.294 4.466−40.975 1.00 20.73 ATOM 577 CA ARG A 76 −17.184 4.551 −41.910 1.00 21.56ATOM 578 CB ARG A 76 −17.544 5.460 −43.101 1.00 22.06 ATOM 579 CG ARG A76 −16.452 5.627 −44.168 1.00 24.00 ATOM 580 CD ARG A 76 −15.586 6.895−43.934 1.00 27.60 ATOM 581 NE ARG A 76 −16.275 8.150 −44.280 1.00 30.93ATOM 582 CZ ARG A 76 −15.778 9.378 −44.082 1.00 32.20 ATOM 583 NH1 ARG A76 −14.572 9.556 −43.535 1.00 30.72 ATOM 584 NH2 ARG A 76 −16.491 10.443−44.437 1.00 32.29 ATOM 585 C ARG A 76 −15.942 5.063 −41.162 1.00 20.85ATOM 586 O ARG A 76 −14.858 4.514 −41.296 1.00 20.93 ATOM 587 N ARG A 77−16.116 6.119 −40.378 1.00 20.17 ATOM 588 CA ARG A 77 −14.990 6.723−39.631 1.00 19.61 ATOM 589 CB ARG A 77 −15.419 8.058 −39.009 1.00 19.11ATOM 590 CG ARG A 77 −15.719 9.106 −40.075 1.00 20.18 ATOM 591 CD ARG A77 −16.379 10.299 −39.459 1.00 22.42 ATOM 592 NE ARG A 77 −16.489 11.411−40.396 1.00 24.06 ATOM 593 CZ ARG A 77 −17.501 11.592 −41.243 1.0027.81 ATOM 594 NH1 ARG A 77 −18.508 10.714 −41.303 1.00 28.01 ATOM 595NH2 ARG A 77 −17.509 12.658 −42.033 1.00 27.19 ATOM 596 C ARG A 77−14.425 5.789 −38.570 1.00 18.77 ATOM 597 O ARG A 77 −13.197 5.685−38.411 1.00 18.64 ATOM 598 N ILE A 78 −15.320 5.117 −37.852 1.00 17.79ATOM 599 CA ILE A 78 −14.931 4.125 −36.857 1.00 18.13 ATOM 600 CB ILE A78 −16.165 3.514 −36.151 1.00 17.88 ATOM 601 CG1 ILE A 78 −16.862 4.564−35.282 1.00 18.78 ATOM 602 CD1 ILE A 78 −18.274 4.154 −34.879 1.0019.16 ATOM 603 CG2 ILE A 78 −15.772 2.279 −35.308 1.00 18.15 ATOM 604 CILE A 78 −14.105 3.012 −37.491 1.00 18.09 ATOM 605 O ILE A 78 −13.0882.612 −36.949 1.00 17.42 ATOM 606 N GLU A 79 −14.565 2.495 −38.631 1.0018.82 ATOM 607 CA GLU A 79 −13.826 1.446 −39.341 1.00 20.22 ATOM 608 CBGLU A 79 −14.587 1.017 −40.609 1.00 20.36 ATOM 609 CG GLU A 79 −15.8110.136 −40.312 1.00 22.29 ATOM 610 CD GLU A 79 −16.633 −0.206 −41.5651.00 22.92 ATOM 611 OE1 GLU A 79 −16.345 0.328 −42.670 1.00 26.99 ATOM612 OE2 GLU A 79 −17.579 −1.012 −41.425 1.00 25.69 ATOM 613 C GLU A 79−12.418 1.918 −39.704 1.00 18.80 ATOM 614 O GLU A 79 −11.450 1.191−39.507 1.00 19.44 ATOM 615 N GLN A 80 −12.301 3.147 −40.211 1.00 18.84ATOM 616 CA GLN A 80 −10.998 3.661 −40.636 1.00 17.90 ATOM 617 CB GLN A80 −11.149 4.921 −41.482 1.00 18.92 ATOM 618 CG GLN A 80 −11.794 4.593−42.844 1.00 21.99 ATOM 619 CD GLN A 80 −12.040 5.799 −43.707 1.00 27.48ATOM 620 OE1 GLN A 80 −12.265 6.898 −43.223 1.00 30.45 ATOM 621 NE2 GLNA 80 −12.037 5.586 −45.013 1.00 32.63 ATOM 622 C GLN A 80 −10.059 3.892−39.456 1.00 17.64 ATOM 623 O GLN A 80 −8.862 3.612 −39.535 1.00 17.21ATOM 624 N TYR A 81 −10.607 4.408 −38.365 1.00 17.29 ATOM 625 CA TYR A81 −9.839 4.552 −37.122 1.00 17.64 ATOM 626 CB TYR A 81 −10.750 5.139−36.023 1.00 17.24 ATOM 627 CG TYR A 81 −10.188 4.973 −34.621 1.00 17.79ATOM 628 CD1 TYR A 81 −9.085 5.728 −34.184 1.00 16.36 ATOM 629 CE1 TYR A81 −8.561 5.568 −32.882 1.00 17.45 ATOM 630 CZ TYR A 81 −9.146 4.646−32.009 1.00 16.35 ATOM 631 OH TYR A 81 −8.654 4.457 −30.724 1.00 17.06ATOM 632 CE2 TYR A 81 −10.238 3.890 −32.423 1.00 17.51 ATOM 633 CD2 TYRA 81 −10.754 4.055 −33.729 1.00 17.00 ATOM 634 C TYR A 81 −9.271 3.197−36.686 1.00 18.04 ATOM 635 O TYR A 81 −8.098 3.083 −36.321 1.00 17.85ATOM 636 N ILE A 82 −10.096 2.159 −36.746 1.00 17.99 ATOM 637 CA ILE A82 −9.661 0.839 −36.295 1.00 19.35 ATOM 638 CB ILE A 82 −10.844 −0.166−36.187 1.00 18.93 ATOM 639 CG1 ILE A 82 −11.753 0.233 −35.017 1.0019.40 ATOM 640 CD1 ILE A 82 −13.093 −0.565 −34.896 1.00 20.46 ATOM 641CG2 ILE A 82 −10.301 −1.587 −35.984 1.00 20.61 ATOM 642 C ILE A 82−8.547 0.292 −37.194 1.00 19.92 ATOM 643 O ILE A 82 −7.543 −0.239−36.708 1.00 20.26 ATOM 644 N THR A 83 −8.713 0.432 −38.503 1.00 20.05ATOM 645 CA THR A 83 −7.709 −0.100 −39.406 1.00 21.11 ATOM 646 CB THR A83 −8.241 −0.297 −40.845 1.00 21.63 ATOM 647 OG1 THR A 83 −8.830 0.902−41.306 1.00 25.88 ATOM 648 CG2 THR A 83 −9.330 −1.347 −40.851 1.0021.56 ATOM 649 C THR A 83 −6.410 0.690 −39.337 1.00 20.59 ATOM 650 O THRA 83 −5.338 0.105 −39.511 1.00 20.72 ATOM 651 N ALA A 84 −6.494 1.997−39.050 1.00 19.51 ATOM 652 CA ALA A 84 −5.292 2.809 −38.777 1.00 19.37ATOM 653 CB ALA A 84 −5.652 4.290 −38.507 1.00 19.42 ATOM 654 C ALA A 84−4.436 2.231 −37.643 1.00 19.32 ATOM 655 O ALA A 84 −3.208 2.370 −37.6491.00 19.47 ATOM 656 N GLN A 85 −5.063 1.535 −36.695 1.00 19.34 ATOM 657CA GLN A 85 −4.325 0.998 −35.544 1.00 18.78 ATOM 658 CB GLN A 85 −5.2660.609 −34.396 1.00 19.29 ATOM 659 CG GLN A 85 −6.260 1.735 −34.007 1.0017.98 ATOM 660 CD GLN A 85 −5.593 3.098 −33.830 1.00 17.20 ATOM 661 OE1GLN A 85 −6.021 4.095 −34.418 1.00 20.82 ATOM 662 NE2 GLN A 85 −4.5403.143 −33.034 1.00 13.47 ATOM 663 C GLN A 85 −3.447 −0.178 −35.932 1.0019.08 ATOM 664 O GLN A 85 −2.478 −0.473 −35.251 1.00 17.94 ATOM 665 NVAL A 86 −3.808 −0.838 −37.032 1.00 19.32 ATOM 666 CA VAL A 86 −2.999−1.928 −37.588 1.00 20.79 ATOM 667 CB VAL A 86 −3.670 −2.581 −38.8231.00 21.18 ATOM 668 CG1 VAL A 86 −2.712 −3.615 −39.454 1.00 22.66 ATOM669 CG2 VAL A 86 −4.980 −3.250 −38.400 1.00 21.47 ATOM 670 C VAL A 86−1.617 −1.381 −37.940 1.00 20.63 ATOM 671 O VAL A 86 −0.602 −1.930−37.520 1.00 21.41 ATOM 672 N THR A 87 −1.604 −0.251 −38.641 1.00 20.67ATOM 673 CA THR A 87 −0.361 0.419 −39.015 1.00 21.10 ATOM 674 CB THR A87 −0.659 1.583 −39.986 1.00 21.59 ATOM 675 OG1 THR A 87 −1.176 1.033−41.202 1.00 23.63 ATOM 676 CG2 THR A 87 0.585 2.370 −40.305 1.00 22.02ATOM 677 C THR A 87 0.412 0.881 −37.795 1.00 20.28 ATOM 678 O THR A 871.620 0.641 −37.679 1.00 20.22 ATOM 679 N LEU A 88 −0.280 1.543 −36.8741.00 19.11 ATOM 680 CA LEU A 88 0.367 2.097 −35.697 1.00 18.71 ATOM 681CB LEU A 88 −0.585 3.015 −34.903 1.00 18.08 ATOM 682 CG LEU A 88 −1.0164.294 −35.596 1.00 18.50 ATOM 683 CD1 LEU A 88 −2.038 5.058 −34.706 1.0019.12 ATOM 684 CD2 LEU A 88 0.206 5.193 −35.937 1.00 19.81 ATOM 685 CLEU A 88 0.976 1.057 −34.780 1.00 18.33 ATOM 686 O LEU A 88 2.101 1.244−34.336 1.00 18.76 ATOM 687 N GLN A 89 0.255 −0.029 −34.492 1.00 18.75ATOM 688 CA GLN A 89 0.809 −1.085 −33.623 1.00 19.67 ATOM 689 CB GLN A89 −0.199 −2.201 −33.373 1.00 19.69 ATOM 690 CG GLN A 89 −1.397 −1.775−32.564 1.00 19.25 ATOM 691 CD GLN A 89 −2.140 −2.951 −32.004 1.00 20.83ATOM 692 OE1 GLN A 89 −2.121 −4.037 −32.580 1.00 19.26 ATOM 693 NE2 GLNA 89 −2.802 −2.751 −30.861 1.00 19.61 ATOM 694 C GLN A 89 2.097 −1.683−34.203 1.00 20.68 ATOM 695 O GLN A 89 3.013 −2.026 −33.461 1.00 21.21ATOM 696 N GLY A 90 2.164 −1.778 −35.524 1.00 21.75 ATOM 697 CA GLY A 903.330 −2.374 −36.173 1.00 23.35 ATOM 698 C GLY A 90 4.604 −1.552 −36.0961.00 24.79 ATOM 699 O GLY A 90 5.699 −2.104 −36.299 1.00 25.52 ATOM 700N ASN A 91 4.477 −0.252 −35.796 1.00 25.17 ATOM 701 CA ASN A 91 5.5960.714 −35.870 1.00 26.14 ATOM 702 CB ASN A 91 5.108 2.161 −35.653 1.0026.83 ATOM 703 CG ASN A 91 4.615 2.849 −36.919 1.00 29.67 ATOM 704 OD1ASN A 91 4.869 2.414 −38.051 1.00 34.53 ATOM 705 ND2 ASN A 91 3.9273.981 −36.724 1.00 32.76 ATOM 706 C ASN A 91 6.656 0.489 −34.820 1.0025.67 ATOM 707 O ASN A 91 6.346 0.385 −33.644 1.00 25.70 ATOM 708 N SERA 92 7.918 0.472 −35.227 1.00 25.32 ATOM 709 CA SER A 92 8.990 0.668−34.257 1.00 25.27 ATOM 710 CB SER A 92 10.314 0.107 −34.775 1.00 26.03ATOM 711 OG SER A 92 10.212 −1.305 −34.803 1.00 30.67 ATOM 712 C SER A92 9.103 2.171 −34.003 1.00 23.80 ATOM 713 O SER A 92 9.055 2.979−34.942 1.00 24.56 ATOM 714 N ASN A 93 9.246 2.544 −32.743 1.00 22.30ATOM 715 CA ASN A 93 9.236 3.953 −32.383 1.00 21.23 ATOM 716 CB ASN A 937.769 4.423 −32.201 1.00 21.41 ATOM 717 CG ASN A 93 7.075 3.704 −31.0511.00 19.23 ATOM 718 OD1 ASN A 93 7.564 3.736 −29.927 1.00 17.81 ATOM 719ND2 ASN A 93 5.974 3.024 −31.335 1.00 19.43 ATOM 720 C ASN A 93 10.1034.149 −31.150 1.00 20.39 ATOM 721 O ASN A 93 10.625 3.154 −30.607 1.0019.46 ATOM 722 N PRO A 94 10.337 5.410 −30.732 1.00 19.95 ATOM 723 CAPRO A 94 11.228 5.604 −29.574 1.00 19.89 ATOM 724 CB PRO A 94 11.2357.131 −29.385 1.00 19.51 ATOM 725 CG PRO A 94 10.988 7.653 −30.753 1.0019.96 ATOM 726 CD PRO A 94 9.952 6.717 −31.325 1.00 19.99 ATOM 727 C PROA 94 10.870 4.898 −28.263 1.00 20.64 ATOM 728 O PRO A 94 11.756 4.727−27.430 1.00 20.54 ATOM 729 N SER A 95 9.610 4.485 −28.073 1.00 20.36ATOM 730 CA SER A 95 9.264 3.674 −26.902 1.00 21.00 ATOM 731 CB SER A 957.770 3.736 −26.587 1.00 20.05 ATOM 732 OG SER A 95 7.413 5.036 −26.1471.00 19.97 ATOM 733 C SER A 95 9.679 2.204 −27.066 1.00 21.73 ATOM 734 OSER A 95 9.809 1.499 −26.072 1.00 22.10 ATOM 735 N GLY A 96 9.853 1.748−28.306 1.00 21.90 ATOM 736 CA GLY A 96 10.229 0.350 −28.562 1.00 23.56ATOM 737 C GLY A 96 9.506 −0.196 −29.774 1.00 24.14 ATOM 738 O GLY A 969.121 0.557 −30.664 1.00 24.24 ATOM 739 N SER A 97 9.315 −1.510 −29.8281.00 25.24 ATOM 740 CA SER A 97 8.703 −2.116 −31.000 1.00 25.77 ATOM 741CB SER A 97 9.751 −2.874 −31.834 1.00 27.15 ATOM 742 OG SER A 97 10.120−4.086 −31.189 1.00 30.57 ATOM 743 C SER A 97 7.590 −3.042 −30.571 1.0025.27 ATOM 744 O SER A 97 7.346 −3.199 −29.376 1.00 24.85 ATOM 745 N LEUA 98 6.930 −3.655 −31.543 1.00 24.82 ATOM 746 CA LEU A 98 5.826 −4.559−31.252 1.00 25.88 ATOM 747 CB LEU A 98 4.982 −4.813 −32.504 1.00 25.31ATOM 748 CG LEU A 98 3.714 −5.673 −32.420 1.00 25.89 ATOM 749 CD1 LEU A98 2.745 −5.169 −31.337 1.00 25.58 ATOM 750 CD2 LEU A 98 3.006 −5.724−33.778 1.00 26.27 ATOM 751 C LEU A 98 6.310 −5.866 −30.604 1.00 26.75ATOM 752 O LEU A 98 5.607 −6.438 −29.762 1.00 27.41 ATOM 753 N ALA A 997.528 −6.290 −30.950 1.00 27.23 ATOM 754 CA ALA A 99 8.074 −7.590−30.533 1.00 27.89 ATOM 755 CB ALA A 99 9.566 −7.700 −30.935 1.00 27.68ATOM 756 C ALA A 99 7.893 −7.911 −29.053 1.00 27.86 ATOM 757 O ALA A 997.450 −9.007 −28.711 1.00 28.77 ATOM 758 N ASP A 100 8.241 −6.966−28.181 1.00 27.75 ATOM 759 CA ASP A 100 8.030 −7.137 −26.741 1.00 27.20ATOM 760 CB ASP A 100 9.328 −6.937 −25.966 1.00 27.10 ATOM 761 CG ASP A100 9.845 −5.525 −26.038 1.00 30.19 ATOM 762 OD1 ASP A 100 10.891 −5.281−25.419 1.00 32.28 ATOM 763 OD2 ASP A 100 9.225 −4.654 −26.694 1.0030.36 ATOM 764 C ASP A 100 6.905 −6.256 −26.173 1.00 25.74 ATOM 765 OASP A 100 6.761 −6.108 −24.956 1.00 26.33 ATOM 766 N GLY A 101 6.118−5.683 −27.075 1.00 24.93 ATOM 767 CA GLY A 101 4.982 −4.853 −26.7071.00 23.22 ATOM 768 C GLY A 101 5.326 −3.418 −26.342 1.00 22.68 ATOM 769O GLY A 101 4.419 −2.580 −26.287 1.00 21.48 ATOM 770 N SER A 102 6.609−3.117 −26.126 1.00 21.53 ATOM 771 CA SER A 102 6.996 −1.815 −25.5631.00 21.49 ATOM 772 CB SER A 102 8.483 −1.739 −25.199 1.00 22.07 ATOM773 OG SER A 102 9.283 −1.958 −26.345 1.00 21.77 ATOM 774 C SER A 1026.604 −0.643 −26.449 1.00 20.66 ATOM 775 O SER A 102 6.279 0.403 −25.9251.00 20.67 ATOM 776 N GLY A 103 6.636 −0.819 −27.771 1.00 19.97 ATOM 777CA GLY A 103 6.257 0.255 −28.707 1.00 19.46 ATOM 778 C GLY A 103 4.8240.777 −28.539 1.00 18.94 ATOM 779 O GLY A 103 4.525 1.903 −28.945 1.0018.13 ATOM 780 N LEU A 104 3.939 −0.043 −27.956 1.00 18.13 ATOM 781 CALEU A 104 2.517 0.326 −27.818 1.00 16.92 ATOM 782 CB LEU A 104 1.672−0.924 −27.447 1.00 17.28 ATOM 783 CG LEU A 104 1.715 −2.104 −28.4301.00 16.76 ATOM 784 CD1 LEU A 104 1.072 −3.356 −27.836 1.00 21.62 ATOM785 CD2 LEU A 104 1.069 −1.751 −29.761 1.00 19.06 ATOM 786 C LEU A 1042.283 1.464 −26.798 1.00 16.57 ATOM 787 O LEU A 104 1.202 2.092 −26.8071.00 16.79 ATOM 788 N GLY A 105 3.279 1.713 −25.936 1.00 14.86 ATOM 789CA GLY A 105 3.255 2.802 −24.938 1.00 15.94 ATOM 790 C GLY A 105 3.5584.199 −25.482 1.00 15.05 ATOM 791 O GLY A 105 3.481 5.179 −24.755 1.0015.93 ATOM 792 N GLU A 106 3.869 4.292 −26.780 1.00 14.63 ATOM 793 CAGLU A 106 4.236 5.548 −27.416 1.00 14.49 ATOM 794 CB GLU A 106 4.7285.250 −28.847 1.00 13.83 ATOM 795 CG GLU A 106 5.215 6.470 −29.678 1.0016.14 ATOM 796 CD GLU A 106 6.479 7.155 −29.139 1.00 18.44 ATOM 797 OE1GLU A 106 6.978 6.817 −28.044 1.00 21.44 ATOM 798 OE2 GLU A 106 6.9728.083 −29.817 1.00 21.04 ATOM 799 C GLU A 106 3.012 6.484 −27.413 1.0014.32 ATOM 800 O GLU A 106 1.928 6.074 −27.828 1.00 15.51 ATOM 801 N PROA 107 3.164 7.706 −26.890 1.00 15.21 ATOM 802 CA PRO A 107 2.025 8.645−26.772 1.00 14.90 ATOM 803 CB PRO A 107 2.598 9.809 −25.951 1.00 15.27ATOM 804 CG PRO A 107 3.833 9.290 −25.310 1.00 17.25 ATOM 805 CD PRO A107 4.385 8.238 −26.243 1.00 14.35 ATOM 806 C PRO A 107 1.468 9.219−28.066 1.00 14.48 ATOM 807 O PRO A 107 0.263 9.371 −28.172 1.00 13.94ATOM 808 N LYS A 108 2.320 9.567 −29.027 1.00 14.24 ATOM 809 CA LYS A108 1.837 10.309 −30.204 1.00 14.51 ATOM 810 CB LYS A 108 1.853 11.828−29.960 1.00 14.26 ATOM 811 CG LYS A 108 3.225 12.524 −30.029 1.00 13.88ATOM 812 CD LYS A 108 3.102 14.033 −29.775 1.00 15.15 ATOM 813 CE LYS A108 4.299 14.820 −30.283 1.00 17.08 ATOM 814 NZ LYS A 108 4.341 16.279−29.836 1.00 15.57 ATOM 815 C LYS A 108 2.608 9.953 −31.454 1.00 14.69ATOM 816 O LYS A 108 3.734 9.442 −31.377 1.00 14.67 ATOM 817 N PHE A 1091.976 10.203 −32.594 1.00 14.54 ATOM 818 CA PHE A 109 2.530 9.860−33.902 1.00 15.20 ATOM 819 CB PHE A 109 1.839 8.594 −34.451 1.00 15.67ATOM 820 CG PHE A 109 1.973 7.407 −33.553 1.00 16.81 ATOM 821 CD1 PHE A109 1.081 7.219 −32.490 1.00 17.93 ATOM 822 CE1 PHE A 109 1.205 6.117−31.649 1.00 21.93 ATOM 823 CZ PHE A 109 2.241 5.210 −31.834 1.00 19.06ATOM 824 CE2 PHE A 109 3.141 5.384 −32.883 1.00 20.27 ATOM 825 CD2 PHE A109 3.003 6.492 −33.737 1.00 19.44 ATOM 826 C PHE A 109 2.301 10.992−34.881 1.00 15.19 ATOM 827 O PHE A 109 1.450 11.861 −34.655 1.00 14.41ATOM 828 N GLU A 110 3.039 10.971 −35.993 1.00 15.14 ATOM 829 CA GLU A110 2.756 11.905 −37.077 1.00 15.24 ATOM 830 CB GLU A 110 3.905 11.933−38.103 1.00 15.00 ATOM 831 CG GLU A 110 5.302 12.204 −37.493 1.00 16.29ATOM 832 CD GLU A 110 5.554 13.673 −37.174 1.00 17.52 ATOM 833 OE1 GLU A110 4.708 14.544 −37.504 1.00 17.65 ATOM 834 OE2 GLU A 110 6.619 13.963−36.587 1.00 18.74 ATOM 835 C GLU A 110 1.462 11.476 −37.762 1.00 15.32ATOM 836 O GLU A 110 1.093 10.278 −37.753 1.00 15.26 ATOM 837 N LEU A111 0.776 12.445 −38.360 1.00 15.59 ATOM 838 CA LEU A 111 −0.522 12.186−39.009 1.00 16.33 ATOM 839 CB LEU A 111 −1.265 13.510 −39.163 1.0016.57 ATOM 840 CG LEU A 111 −1.770 13.908 −37.756 1.00 18.11 ATOM 841CD1 LEU A 111 −1.819 15.405 −37.574 1.00 21.43 ATOM 842 CD2 LEU A 111−3.151 13.204 −37.506 1.00 18.74 ATOM 843 C LEU A 111 −0.409 11.436−40.350 1.00 17.37 ATOM 844 O LEU A 111 −1.426 11.023 −40.944 1.00 17.23ATOM 845 N THR A 112 0.833 11.258 −40.815 1.00 17.42 ATOM 846 CA THR A112 1.144 10.301 −41.887 1.00 17.92 ATOM 847 CB THR A 112 2.512 10.619−42.499 1.00 17.98 ATOM 848 OG1 THR A 112 3.476 10.702 −41.445 1.0018.32 ATOM 849 CG2 THR A 112 2.486 11.945 −43.228 1.00 19.47 ATOM 850 CTHR A 112 1.215 8.846 −41.356 1.00 18.90 ATOM 851 O THR A 112 1.5357.917 −42.117 1.00 17.68 ATOM 852 N LEU A 113 0.944 8.664 −40.055 1.0018.48 ATOM 853 CA LEU A 113 1.041 7.379 −39.348 1.00 19.78 ATOM 854 CBLEU A 113 0.061 6.319 −39.904 1.00 19.45 ATOM 855 CG LEU A 113 −1.4116.699 −40.074 1.00 21.71 ATOM 856 CD1 LEU A 113 −2.194 5.470 −40.4771.00 23.46 ATOM 857 CD2 LEU A 113 −2.005 7.323 −38.800 1.00 21.59 ATOM858 C LEU A 113 2.481 6.866 −39.338 1.00 20.43 ATOM 859 O LEU A 1132.737 5.704 −39.653 1.00 20.96 ATOM 860 N LYS A 114 3.406 7.769 −39.0241.00 20.07 ATOM 861 CA LYS A 114 4.826 7.460 −38.863 1.00 20.25 ATOM 862CB LYS A 114 5.662 8.209 −39.899 1.00 20.95 ATOM 863 CG LYS A 114 5.4327.725 −41.314 1.00 25.23 ATOM 864 CD LYS A 114 6.636 8.059 −42.184 1.0033.76 ATOM 865 CE LYS A 114 6.551 7.360 −43.537 1.00 38.31 ATOM 866 NZLYS A 114 5.285 7.711 −44.251 1.00 41.29 ATOM 867 C LYS A 114 5.2527.874 −37.471 1.00 19.64 ATOM 868 O LYS A 114 4.576 8.708 −36.845 1.0019.39 ATOM 869 N PRO A 115 6.376 7.318 −36.973 1.00 19.06 ATOM 870 CAPRO A 115 6.750 7.626 −35.601 1.00 18.72 ATOM 871 CB PRO A 115 7.9636.712 −35.326 1.00 19.82 ATOM 872 CG PRO A 115 8.101 5.814 −36.500 1.0020.93 ATOM 873 CD PRO A 115 7.339 6.412 −37.639 1.00 19.08 ATOM 874 CPRO A 115 7.156 9.093 −35.434 1.00 18.70 ATOM 875 O PRO A 115 7.6949.724 −36.375 1.00 17.25 ATOM 876 N PHE A 116 6.844 9.628 −34.256 1.0018.55 ATOM 877 CA PHE A 116 7.342 10.918 −33.805 1.00 18.27 ATOM 878 CBPHE A 116 6.359 11.566 −32.809 1.00 18.41 ATOM 879 CG PHE A 116 6.90812.804 −32.151 1.00 17.33 ATOM 880 CD1 PHE A 116 6.942 14.014 −32.8471.00 16.98 ATOM 881 CE1 PHE A 116 7.457 15.173 −32.254 1.00 16.22 ATOM882 CZ PHE A 116 7.950 15.138 −30.935 1.00 15.88 ATOM 883 CE2 PHE A 1167.902 13.917 −30.216 1.00 16.74 ATOM 884 CD2 PHE A 116 7.380 12.767−30.825 1.00 16.05 ATOM 885 C PHE A 116 8.701 10.695 −33.141 1.00 19.14ATOM 886 O PHE A 116 8.808 9.987 −32.134 1.00 19.66 ATOM 887 N THR A 1179.746 11.299 −33.713 1.00 19.53 ATOM 888 CA THR A 117 11.116 11.020−33.315 1.00 20.15 ATOM 889 CB THR A 117 12.042 10.999 −34.567 1.0020.83 ATOM 890 OG1 THR A 117 11.988 12.277 −35.222 1.00 22.59 ATOM 891CG2 THR A 117 11.576 9.895 −35.557 1.00 21.40 ATOM 892 C THR A 11711.685 11.969 −32.263 1.00 20.34 ATOM 893 O THR A 117 12.813 11.768−31.774 1.00 21.13 ATOM 894 N GLY A 118 10.943 13.017 −31.914 1.00 19.25ATOM 895 CA GLY A 118 11.451 14.018 −30.974 1.00 19.41 ATOM 896 C GLY A118 11.431 13.498 −29.541 1.00 19.75 ATOM 897 O GLY A 118 10.913 12.397−29.281 1.00 19.99 ATOM 898 N ASN A 119 11.998 14.279 −28.622 1.00 19.56ATOM 899 CA ASN A 119 11.958 13.954 −27.198 1.00 20.41 ATOM 900 CB ASN A119 12.961 14.801 −26.419 1.00 21.50 ATOM 901 CG ASN A 119 14.377 14.612−26.930 1.00 25.14 ATOM 902 OD1 ASN A 119 14.779 13.500 −27.294 1.0030.77 ATOM 903 ND2 ASN A 119 15.131 15.693 −26.987 1.00 31.23 ATOM 904 CASN A 119 10.550 14.194 −26.696 1.00 20.06 ATOM 905 O ASN A 119 9.88115.089 −27.167 1.00 19.00 ATOM 906 N TRP A 120 10.084 13.348 −25.7871.00 19.74 ATOM 907 CA TRP A 120 8.707 13.466 −25.316 1.00 19.03 ATOM908 CB TRP A 120 7.717 12.917 −26.359 1.00 18.71 ATOM 909 CG TRP A 1206.351 13.522 −26.162 1.00 19.76 ATOM 910 CD1 TRP A 120 5.239 12.901−25.673 1.00 19.81 ATOM 911 NE1 TRP A 120 4.186 13.799 −25.593 1.0019.49 ATOM 912 CE2 TRP A 120 4.612 15.021 −26.042 1.00 19.44 ATOM 913CD2 TRP A 120 5.975 14.886 −26.410 1.00 19.35 ATOM 914 CE3 TRP A 1206.657 16.014 −26.895 1.00 18.94 ATOM 915 CZ3 TRP A 120 5.959 17.220−27.010 1.00 20.22 ATOM 916 CH2 TRP A 120 4.602 17.315 −26.628 1.0020.15 ATOM 917 CZ2 TRP A 120 3.918 16.233 −26.160 1.00 18.83 ATOM 918 CTRP A 120 8.602 12.685 −24.001 1.00 18.80 ATOM 919 O TRP A 120 9.45411.833 −23.722 1.00 18.83 ATOM 920 N GLY A 121 7.593 12.990 −23.189 1.0017.91 ATOM 921 CA GLY A 121 7.314 12.189 −21.988 1.00 17.64 ATOM 922 CGLY A 121 6.721 10.834 −22.362 1.00 18.93 ATOM 923 O GLY A 121 5.49910.704 −22.487 1.00 18.99 ATOM 924 N ARG A 122 7.589 9.828 −22.536 1.0017.95 ATOM 925 CA ARG A 122 7.195 8.483 −22.958 1.00 17.86 ATOM 926 CBARG A 122 7.686 8.193 −24.394 1.00 17.37 ATOM 927 CG ARG A 122 9.1818.529 −24.626 1.00 19.53 ATOM 928 CD ARG A 122 9.689 7.987 −25.969 1.0017.88 ATOM 929 NE ARG A 122 9.012 8.549 −27.159 1.00 18.08 ATOM 930 CZARG A 122 9.425 9.645 −27.807 1.00 18.06 ATOM 931 NH1 ARG A 122 10.47710.326 −27.366 1.00 16.88 ATOM 932 NH2 ARG A 122 8.784 10.074 −28.8921.00 17.35 ATOM 933 C ARG A 122 7.799 7.450 −21.976 1.00 17.48 ATOM 934O ARG A 122 8.848 7.697 −21.396 1.00 17.69 ATOM 935 N PRO A 123 7.1426.298 −21.781 1.00 17.14 ATOM 936 CA PRO A 123 5.886 5.916 −22.382 1.0016.01 ATOM 937 CB PRO A 123 5.908 4.385 −22.266 1.00 16.51 ATOM 938 CGPRO A 123 6.585 4.144 −20.969 1.00 16.30 ATOM 939 CD PRO A 123 7.6585.238 −20.873 1.00 16.95 ATOM 940 C PRO A 123 4.716 6.494 −21.581 1.0015.29 ATOM 941 O PRO A 123 4.926 7.057 −20.521 1.00 15.07 ATOM 942 N GLNA 124 3.504 6.362 −22.120 1.00 14.60 ATOM 943 CA GLN A 124 2.289 6.675−21.386 1.00 14.40 ATOM 944 CB GLN A 124 1.602 7.889 −22.001 1.00 14.71ATOM 945 CG GLN A 124 2.442 9.186 −21.711 1.00 11.93 ATOM 946 CD GLN A124 1.993 10.407 −22.472 1.00 15.67 ATOM 947 OE1 GLN A 124 2.807 11.310−22.758 1.00 15.75 ATOM 948 NE2 GLN A 124 0.718 10.450 −22.822 1.00 9.85ATOM 949 C GLN A 124 1.441 5.421 −21.511 1.00 14.28 ATOM 950 O GLN A 1240.988 5.095 −22.604 1.00 14.63 ATOM 951 N ARG A 125 1.241 4.731 −20.3901.00 13.50 ATOM 952 CA ARG A 125 0.700 3.382 −20.398 1.00 13.96 ATOM 953CB ARG A 125 1.331 2.567 −19.256 1.00 14.28 ATOM 954 CG ARG A 125 2.8642.703 −19.249 1.00 15.16 ATOM 955 CD ARG A 125 3.503 1.577 −18.439 1.0018.07 ATOM 956 NE ARG A 125 4.924 1.827 −18.132 1.00 17.46 ATOM 957 CZARG A 125 5.944 1.334 −18.833 1.00 19.43 ATOM 958 NH1 ARG A 125 5.7280.601 −19.925 1.00 20.02 ATOM 959 NH2 ARG A 125 7.197 1.596 −18.453 1.0018.94 ATOM 960 C ARG A 125 −0.829 3.335 −20.359 1.00 14.48 ATOM 961 OARG A 125 −1.424 2.262 −20.343 1.00 14.40 ATOM 962 N ASP A 126 −1.4624.509 −20.374 1.00 14.03 ATOM 963 CA ASP A 126 −2.919 4.568 −20.542 1.0013.39 ATOM 964 CB ASP A 126 −3.488 5.922 −20.067 1.00 13.17 ATOM 965 CGASP A 126 −2.926 7.092 −20.845 1.00 14.17 ATOM 966 OD1 ASP A 126 −1.7137.108 −21.143 1.00 12.29 ATOM 967 OD2 ASP A 126 −3.705 8.003 −21.1871.00 16.69 ATOM 968 C ASP A 126 −3.350 4.306 −21.974 1.00 13.60 ATOM 969O ASP A 126 −4.452 3.806 −22.189 1.00 13.56 ATOM 970 N GLY A 127 −2.4914.634 −22.948 1.00 12.78 ATOM 971 CA GLY A 127 −2.886 4.558 −24.358 1.0013.20 ATOM 972 C GLY A 127 −3.473 3.213 −24.794 1.00 12.77 ATOM 973 OGLY A 127 −4.579 3.150 −25.327 1.00 12.56 ATOM 974 N PRO A 128 −2.7202.120 −24.613 1.00 13.94 ATOM 975 CA PRO A 128 −3.262 0.810 −24.978 1.0012.97 ATOM 976 CB PRO A 128 −2.135 −0.162 −24.552 1.00 14.15 ATOM 977 CGPRO A 128 −0.907 0.656 −24.721 1.00 14.39 ATOM 978 CD PRO A 128 −1.3232.010 −24.150 1.00 13.67 ATOM 979 C PRO A 128 −4.571 0.455 −24.255 1.0013.30 ATOM 980 O PRO A 128 −5.433 −0.161 −24.869 1.00 13.72 ATOM 981 NALA A 129 −4.718 0.852 −22.985 1.00 12.95 ATOM 982 CA ALA A 129 −5.9630.611 −22.258 1.00 13.49 ATOM 983 CB ALA A 129 −5.806 1.016 −20.808 1.0013.00 ATOM 984 C ALA A 129 −7.162 1.329 −22.923 1.00 13.43 ATOM 985 OALA A 129 −8.217 0.721 −23.159 1.00 13.26 ATOM 986 N LEU A 130 −6.9982.619 −23.221 1.00 12.51 ATOM 987 CA LEU A 130 −8.068 3.409 −23.813 1.0012.69 ATOM 988 CB LEU A 130 −7.678 4.903 −23.806 1.00 12.56 ATOM 989 CGLEU A 130 −7.458 5.555 −22.426 1.00 14.76 ATOM 990 CD1 LEU A 130 −6.9596.991 −22.643 1.00 15.23 ATOM 991 CD2 LEU A 130 −8.776 5.544 −21.6511.00 15.30 ATOM 992 C LEU A 130 −8.410 2.934 −25.228 1.00 12.78 ATOM 993O LEU A 130 −9.571 2.863 −25.607 1.00 12.83 ATOM 994 N ARG A 131 −7.3862.601 −26.015 1.00 13.70 ATOM 995 CA ARG A 131 −7.630 2.070 −27.351 1.0014.52 ATOM 996 CB ARG A 131 −6.316 1.925 −28.135 1.00 14.04 ATOM 997 CGARG A 131 −6.550 1.438 −29.566 1.00 15.60 ATOM 998 CD ARG A 131 −5.2781.522 −30.428 1.00 15.59 ATOM 999 NE ARG A 131 −4.118 0.915 −29.779 1.0016.77 ATOM 1000 CZ ARG A 131 −2.860 1.205 −30.098 1.00 16.26 ATOM 1001NH1 ARG A 131 −2.610 2.104 −31.054 1.00 16.37 ATOM 1002 NH2 ARG A 131−1.856 0.618 −29.448 1.00 15.96 ATOM 1003 C ARG A 131 −8.408 0.729−27.283 1.00 14.44 ATOM 1004 O ARG A 131 −9.350 0.533 −28.050 1.00 15.71ATOM 1005 N ALA A 132 −8.025 −0.164 −26.364 1.00 14.50 ATOM 1006 CA ALAA 132 −8.738 −1.456 −26.195 1.00 15.06 ATOM 1007 CB ALA A 132 −8.069−2.348 −25.112 1.00 15.02 ATOM 1008 C ALA A 132 −10.194 −1.197 −25.8461.00 15.43 ATOM 1009 O ALA A 132 −11.101 −1.808 −26.416 1.00 15.57 ATOM1010 N ILE A 133 −10.418 −0.270 −24.915 1.00 15.70 ATOM 1011 CA ILE A133 −11.777 0.049 −24.491 1.00 14.44 ATOM 1012 CB ILE A 133 −11.7751.056 −23.335 1.00 14.07 ATOM 1013 CG1 ILE A 133 −11.268 0.387 −22.0461.00 15.40 ATOM 1014 CD1 ILE A 133 −10.751 1.388 −21.017 1.00 16.66 ATOM1015 CG2 ILE A 133 −13.176 1.702 −23.101 1.00 13.98 ATOM 1016 C ILE A133 −12.633 0.517 −25.679 1.00 14.36 ATOM 1017 O ILE A 133 −13.781 0.102−25.807 1.00 14.69 ATOM 1018 N ALA A 134 −12.079 1.362 −26.545 1.0013.69 ATOM 1019 CA ALA A 134 −12.819 1.832 −27.720 1.00 13.81 ATOM 1020CB ALA A 134 −12.019 2.949 −28.452 1.00 13.99 ATOM 1021 C ALA A 134−13.140 0.662 −28.657 1.00 14.62 ATOM 1022 O ALA A 134 −14.279 0.473−29.087 1.00 14.90 ATOM 1023 N LEU A 135 −12.133 −0.155 −28.947 1.0014.66 ATOM 1024 CA LEU A 135 −12.328 −1.251 −29.901 1.00 15.75 ATOM 1025CB LEU A 135 −10.984 −1.875 −30.311 1.00 15.75 ATOM 1026 CG LEU A 135−10.348 −1.231 −31.557 1.00 16.40 ATOM 1027 CD1 LEU A 135 −10.257 0.308−31.471 1.00 18.56 ATOM 1028 CD2 LEU A 135 −8.980 −1.852 −31.804 1.0017.85 ATOM 1029 C LEU A 135 −13.277 −2.306 −29.340 1.00 15.13 ATOM 1030O LEU A 135 −14.079 −2.845 −30.087 1.00 15.84 ATOM 1031 N ILE A 136−13.192 −2.573 −28.039 1.00 15.22 ATOM 1032 CA ILE A 136 −14.153 −3.473−27.377 1.00 15.71 ATOM 1033 CB ILE A 136 −13.734 −3.829 −25.918 1.0015.92 ATOM 1034 CG1 ILE A 136 −12.408 −4.598 −25.904 1.00 15.39 ATOM1035 CD1 ILE A 136 −11.742 −4.679 −24.497 1.00 15.63 ATOM 1036 CG2 ILE A136 −14.842 −4.611 −25.204 1.00 16.20 ATOM 1037 C ILE A 136 −15.565−2.902 −27.457 1.00 17.10 ATOM 1038 O ILE A 136 −16.531 −3.631 −27.7281.00 17.48 ATOM 1039 N GLY A 137 −15.685 −1.581 −27.297 1.00 16.46 ATOM1040 CA GLY A 137 −16.979 −0.902 −27.484 1.00 16.59 ATOM 1041 C GLY A137 −17.600 −1.206 −28.833 1.00 17.13 ATOM 1042 O GLY A 137 −18.778−1.605 −28.920 1.00 16.92 ATOM 1043 N TYR A 138 −16.817 −1.056 −29.8981.00 16.61 ATOM 1044 CA TYR A 138 −17.353 −1.349 −31.224 1.00 17.93 ATOM1045 CB TYR A 138 −16.446 −0.838 −32.341 1.00 17.46 ATOM 1046 CG TYR A138 −17.112 −0.897 −33.693 1.00 18.28 ATOM 1047 CD1 TYR A 138 −18.350−0.268 −33.914 1.00 18.85 ATOM 1048 CE1 TYR A 138 −18.966 −0.314 −35.1531.00 21.29 ATOM 1049 CZ TYR A 138 −18.358 −1.001 −36.207 1.00 20.99 ATOM1050 OH TYR A 138 −18.994 −1.055 −37.433 1.00 20.06 ATOM 1051 CE2 TYR A138 −17.133 −1.636 −36.026 1.00 19.88 ATOM 1052 CD2 TYR A 138 −16.512−1.583 −34.766 1.00 18.25 ATOM 1053 C TYR A 138 −17.643 −2.844 −31.4061.00 18.67 ATOM 1054 O TYR A 138 −18.654 −3.207 −32.037 1.00 20.06 ATOM1055 N SER A 139 −16.766 −3.686 −30.864 1.00 19.48 ATOM 1056 CA SER A139 −16.942 −5.146 −30.900 1.00 21.02 ATOM 1057 CB SER A 139 −15.808−5.829 −30.129 1.00 21.09 ATOM 1058 OG SER A 139 −14.581 −5.598 −30.7891.00 21.49 ATOM 1059 C SER A 139 −18.298 −5.557 −30.325 1.00 22.24 ATOM1060 O SER A 139 −19.002 −6.392 −30.907 1.00 23.64 ATOM 1061 N LYS A 140−18.669 −4.976 −29.188 1.00 22.77 ATOM 1062 CA LYS A 140 −19.987 −5.225−28.595 1.00 24.29 ATOM 1063 CB LYS A 140 −20.218 −4.343 −27.370 1.0024.29 ATOM 1064 CG LYS A 140 −19.384 −4.695 −26.170 1.00 26.62 ATOM 1065CD LYS A 140 −19.696 −3.693 −25.060 1.00 28.24 ATOM 1066 CE LYS A 140−18.589 −3.635 −24.056 1.00 28.19 ATOM 1067 NZ LYS A 140 −18.940 −2.725−22.954 1.00 26.78 ATOM 1068 C LYS A 140 −21.126 −5.001 −29.584 1.0024.17 ATOM 1069 O LYS A 140 −22.053 −5.823 −29.670 1.00 24.94 ATOM 1070N TRP A 141 −21.062 −3.898 −30.321 1.00 23.60 ATOM 1071 CA TRP A 141−22.054 −3.613 −31.338 1.00 24.29 ATOM 1072 CB TRP A 141 −21.847 −2.226−31.953 1.00 24.36 ATOM 1073 CG TRP A 141 −22.973 −1.833 −32.874 1.0024.25 ATOM 1074 CD1 TRP A 141 −24.113 −1.170 −32.531 1.00 25.06 ATOM1075 NE1 TRP A 141 −24.921 −1.016 −33.638 1.00 25.22 ATOM 1076 CE2 TRP A141 −24.302 −1.575 −34.722 1.00 24.07 ATOM 1077 CD2 TRP A 141 −23.078−2.115 −34.276 1.00 24.80 ATOM 1078 CE3 TRP A 141 −22.248 −2.766 −35.2031.00 25.40 ATOM 1079 CZ3 TRP A 141 −22.669 −2.858 −36.532 1.00 25.72ATOM 1080 CH2 TRP A 141 −23.891 −2.304 −36.940 1.00 24.95 ATOM 1081 CZ2TRP A 141 −24.721 −1.666 −36.051 1.00 25.14 ATOM 1082 C TRP A 141−22.078 −4.666 −32.448 1.00 24.47 ATOM 1083 O TRP A 141 −23.155 −5.152−32.831 1.00 24.52 ATOM 1084 N LEU A 142 −20.904 −4.991 −32.985 1.0024.39 ATOM 1085 CA LEU A 142 −20.806 −6.024 −34.010 1.00 25.05 ATOM 1086CB LEU A 142 −19.361 −6.199 −34.473 1.00 24.56 ATOM 1087 CG LEU A 142−18.754 −5.023 −35.252 1.00 24.44 ATOM 1088 CD1 LEU A 142 −17.274 −5.304−35.441 1.00 23.72 ATOM 1089 CD2 LEU A 142 −19.424 −4.793 −36.624 1.0025.76 ATOM 1090 C LEU A 142 −21.406 −7.364 −33.556 1.00 25.97 ATOM 1091O LEU A 142 −22.195 −7.966 −34.283 1.00 26.49 ATOM 1092 N ILE A 143−21.045 −7.814 −32.359 1.00 27.04 ATOM 1093 CA ILE A 143 −21.596 −9.040−31.792 1.00 28.46 ATOM 1094 CB ILE A 143 −20.959 −9.362 −30.425 1.0028.30 ATOM 1095 CG1 ILE A 143 −19.474 −9.722 −30.609 1.00 27.97 ATOM1096 CD1 ILE A 143 −18.707 −9.814 −29.301 1.00 29.85 ATOM 1097 CG2 ILE A143 −21.720 −10.494 −29.717 1.00 29.57 ATOM 1098 C ILE A 143 −23.124−8.992 −31.682 1.00 29.52 ATOM 1099 O ILE A 143 −23.813 −9.928 −32.1181.00 30.10 ATOM 1100 N ASN A 144 −23.655 −7.916 −31.111 1.00 30.37 ATOM1101 CA ASN A 144 −25.109 −7.768 −30.988 1.00 32.18 ATOM 1102 CB ASN A144 −25.479 −6.522 −30.186 1.00 32.73 ATOM 1103 CG ASN A 144 −26.960−6.489 −29.792 1.00 36.88 ATOM 1104 OD1 ASN A 144 −27.444 −7.350 −29.0411.00 42.25 ATOM 1105 ND2 ASN A 144 −27.685 −5.488 −30.291 1.00 40.10ATOM 1106 C ASN A 144 −25.820 −7.760 −32.341 1.00 32.17 ATOM 1107 O ASNA 144 −27.012 −8.029 −32.411 1.00 32.73 ATOM 1108 N ASN A 145 −25.094−7.460 −33.411 1.00 32.19 ATOM 1109 CA ASN A 145 −25.705 −7.403 −34.7261.00 32.78 ATOM 1110 CB ASN A 145 −25.526 −6.014 −35.331 1.00 33.16 ATOM1111 CG ASN A 145 −26.397 −4.986 −34.639 1.00 34.32 ATOM 1112 OD1 ASN A145 −27.576 −4.841 −34.969 1.00 37.42 ATOM 1113 ND2 ASN A 145 −25.834−4.289 −33.647 1.00 34.31 ATOM 1114 C ASN A 145 −25.285 −8.533 −35.6711.00 33.13 ATOM 1115 O ASN A 145 −25.412 −8.415 −36.902 1.00 33.03 ATOM1116 N ASN A 146 −24.789 −9.618 −35.065 1.00 33.43 ATOM 1117 CA ASN A146 −24.475 −10.885 −35.736 1.00 34.14 ATOM 1118 CB ASN A 146 −25.710−11.459 −36.461 1.00 34.85 ATOM 1119 CG ASN A 146 −26.994 −11.280−35.657 1.00 37.41 ATOM 1120 OD1 ASN A 146 −27.033 −11.543 −34.450 1.0041.43 ATOM 1121 ND2 ASN A 146 −28.047 −10.814 −36.321 1.00 41.54 ATOM1122 C ASN A 146 −23.266 −10.795 −36.652 1.00 33.92 ATOM 1123 O ASN A146 −23.216 −11.419 −37.724 1.00 33.76 ATOM 1124 N TYR A 147 −22.280−10.013 −36.221 1.00 32.92 ATOM 1125 CA TYR A 147 −21.049 −9.870 −36.9741.00 32.90 ATOM 1126 CB TYR A 147 −20.859 −8.423 −37.451 1.00 33.31 ATOM1127 CG TYR A 147 −21.966 −7.893 −38.339 1.00 33.29 ATOM 1128 CD1 TYR A147 −22.168 −8.410 −39.621 1.00 34.11 ATOM 1129 CE1 TYR A 147 −23.177−7.924 −40.438 1.00 34.19 ATOM 1130 CZ TYR A 147 −23.986 −6.888 −39.9831.00 34.10 ATOM 1131 OH TYR A 147 −24.987 −6.399 −40.794 1.00 35.08 ATOM1132 CE2 TYR A 147 −23.798 −6.345 −38.722 1.00 32.48 ATOM 1133 CD2 TYR A147 −22.792 −6.852 −37.906 1.00 32.03 ATOM 1134 C TYR A 147 −19.857−10.297 −36.138 1.00 32.74 ATOM 1135 O TYR A 147 −18.795 −9.710 −36.2421.00 32.03 ATOM 1136 N GLN A 148 −20.037 −11.325 −35.312 1.00 33.44 ATOM1137 CA GLN A 148 −18.977 −11.807 −34.427 1.00 34.39 ATOM 1138 CB GLN A148 −19.483 −12.971 −33.573 1.00 34.76 ATOM 1139 CG GLN A 148 −18.523−13.445 −32.481 1.00 35.56 ATOM 1140 CD GLN A 148 −19.216 −14.273−31.401 1.00 36.59 ATOM 1141 OE1 GLN A 148 −20.296 −13.913 −30.916 1.0041.20 ATOM 1142 NE2 GLN A 148 −18.589 −15.380 −31.008 1.00 38.65 ATOM1143 C GLN A 148 −17.690 −12.196 −35.176 1.00 34.46 ATOM 1144 O GLN A148 −16.582 −12.002 −34.654 1.00 34.20 ATOM 1145 N PHE A 149 −17.841−12.735 −36.391 1.00 34.19 ATOM 1146 CA PHE A 149 −16.696 −13.131−37.217 1.00 34.10 ATOM 1147 CB PHE A 149 −17.140 −13.804 −38.534 1.0035.60 ATOM 1148 CG PHE A 149 −18.346 −13.168 −39.193 1.00 38.70 ATOM1149 CD1 PHE A 149 −19.388 −13.976 −39.676 1.00 42.72 ATOM 1150 CE1 PHEA 149 −20.518 −13.417 −40.295 1.00 43.69 ATOM 1151 CZ PHE A 149 −20.615−12.019 −40.437 1.00 43.12 ATOM 1152 CE2 PHE A 149 −19.567 −11.188−39.953 1.00 43.20 ATOM 1153 CD2 PHE A 149 −18.451 −11.772 −39.341 1.0042.33 ATOM 1154 C PHE A 149 −15.746 −11.960 −37.517 1.00 32.49 ATOM 1155O PHE A 149 −14.528 −12.132 −37.548 1.00 32.23 ATOM 1156 N THR A 150−16.327 −10.789 −37.751 1.00 30.95 ATOM 1157 CA THR A 150 −15.570 −9.568−38.040 1.00 29.68 ATOM 1158 CB THR A 150 −16.512 −8.424 −38.445 1.0029.97 ATOM 1159 OG1 THR A 150 −17.162 −8.768 −39.673 1.00 30.58 ATOM1160 CG2 THR A 150 −15.758 −7.096 −38.637 1.00 29.18 ATOM 1161 C THR A150 −14.727 −9.203 −36.822 1.00 28.54 ATOM 1162 O THR A 150 −13.566−8.827 −36.965 1.00 28.38 ATOM 1163 N VAL A 151 −15.310 −9.354 −35.6361.00 27.23 ATOM 1164 CA VAL A 151 −14.597 −9.146 −34.374 1.00 26.78 ATOM1165 CB VAL A 151 −15.529 −9.352 −33.148 1.00 26.25 ATOM 1166 CG1 VAL A151 −14.752 −9.213 −31.832 1.00 26.50 ATOM 1167 CG2 VAL A 151 −16.690−8.361 −33.178 1.00 24.41 ATOM 1168 C VAL A 151 −13.384 −10.080 −34.3051.00 27.61 ATOM 1169 O VAL A 151 −12.246 −9.638 −34.106 1.00 26.67 ATOM1170 N SER A 152 −13.625 −11.375 −34.505 1.00 28.53 ATOM 1171 CA SER A152 −12.551 −12.369 −34.470 1.00 30.01 ATOM 1172 CB SER A 152 −13.102−13.759 −34.773 1.00 30.18 ATOM 1173 OG SER A 152 −13.612 −14.300−33.586 1.00 32.28 ATOM 1174 C SER A 152 −11.419 −12.091 −35.430 1.0030.23 ATOM 1175 O SER A 152 −10.250 −12.250 −35.090 1.00 30.95 ATOM 1176N ASN A 153 −11.762 −11.705 −36.641 1.00 31.46 ATOM 1177 CA ASN A 153−10.753 −11.624 −37.674 1.00 32.41 ATOM 1178 CB ASN A 153 −11.333−12.118 −38.997 1.00 33.62 ATOM 1179 CG ASN A 153 −11.791 −13.584−38.902 1.00 36.13 ATOM 1180 OD1 ASN A 153 −12.931 −13.918 −39.231 1.0040.99 ATOM 1181 ND2 ASN A 153 −10.917 −14.444 −38.383 1.00 37.61 ATOM1182 C ASN A 153 −10.060 −10.272 −37.787 1.00 32.10 ATOM 1183 O ASN A153 −8.850 −10.213 −38.020 1.00 32.79 ATOM 1184 N VAL A 154 −10.810−9.193 −37.577 1.00 30.55 ATOM 1185 CA VAL A 154 −10.251 −7.854 −37.7501.00 28.93 ATOM 1186 CB VAL A 154 −11.217 −6.925 −38.537 1.00 29.03 ATOM1187 CG1 VAL A 154 −10.565 −5.577 −38.827 1.00 29.23 ATOM 1188 CG2 VAL A154 −11.654 −7.585 −39.860 1.00 29.87 ATOM 1189 C VAL A 154 −9.824−7.211 −36.414 1.00 27.28 ATOM 1190 O VAL A 154 −8.722 −6.678 −36.3061.00 26.96 ATOM 1191 N ILE A 155 −10.685 −7.288 −35.403 1.00 25.13 ATOM1192 CA ILE A 155 −10.525 −6.459 −34.197 1.00 23.05 ATOM 1193 CB ILE A155 −11.900 −5.972 −33.670 1.00 23.10 ATOM 1194 CG1 ILE A 155 −12.596−5.128 −34.741 1.00 22.49 ATOM 1195 CD1 ILE A 155 −14.006 −4.680 −34.3751.00 22.72 ATOM 1196 CG2 ILE A 155 −11.731 −5.144 −32.399 1.00 23.05ATOM 1197 C ILE A 155 −9.710 −7.136 −33.092 1.00 22.44 ATOM 1198 O ILE A155 −8.789 −6.537 −32.533 1.00 21.14 ATOM 1199 N TRP A 156 −10.006−8.409 −32.822 1.00 21.48 ATOM 1200 CA TRP A 156 −9.392 −9.099 −31.6961.00 21.75 ATOM 1201 CB TRP A 156 −9.958 −10.520 −31.511 1.00 22.50 ATOM1202 CG TRP A 156 −9.298 −11.245 −30.371 1.00 23.43 ATOM 1203 CD1 TRP A156 −8.420 −12.298 −30.461 1.00 24.92 ATOM 1204 NE1 TRP A 156 −8.011−12.680 −29.198 1.00 24.85 ATOM 1205 CE2 TRP A 156 −8.600 −11.863−28.269 1.00 26.38 ATOM 1206 CD2 TRP A 156 −9.416 −10.941 −28.970 1.0025.03 ATOM 1207 CE3 TRP A 156 −10.139 −9.983 −28.236 1.00 25.01 ATOM1208 CZ3 TRP A 156 −10.024 −9.982 −26.844 1.00 24.30 ATOM 1209 CH2 TRP A156 −9.206 −10.910 −26.185 1.00 23.97 ATOM 1210 CZ2 TRP A 156 −8.495−11.861 −26.875 1.00 24.60 ATOM 1211 C TRP A 156 −7.845 −9.109 −31.6991.00 21.52 ATOM 1212 O TRP A 156 −7.235 −8.945 −30.648 1.00 21.78 ATOM1213 N PRO A 157 −7.209 −9.303 −32.870 1.00 21.66 ATOM 1214 CA PRO A 157−5.726 −9.258 −32.878 1.00 21.40 ATOM 1215 CB PRO A 157 −5.378 −9.459−34.360 1.00 21.26 ATOM 1216 CG PRO A 157 −6.583 −10.172 −34.955 1.0022.82 ATOM 1217 CD PRO A 157 −7.762 −9.596 −34.207 1.00 21.45 ATOM 1218C PRO A 157 −5.162 −7.898 −32.410 1.00 21.30 ATOM 1219 O PRO A 157−4.092 −7.837 −31.795 1.00 21.11 ATOM 1220 N ILE A 158 −5.881 −6.821−32.724 1.00 20.52 ATOM 1221 CA ILE A 158 −5.457 −5.467 −32.318 1.0019.93 ATOM 1222 CB ILE A 158 −6.273 −4.348 −33.034 1.00 19.73 ATOM 1223CG1 ILE A 158 −6.261 −4.527 −34.559 1.00 21.25 ATOM 1224 CD1 ILE A 158−7.229 −3.640 −35.351 1.00 20.03 ATOM 1225 CG2 ILE A 158 −5.686 −2.971−32.670 1.00 20.16 ATOM 1226 C ILE A 158 −5.632 −5.366 −30.816 1.0019.58 ATOM 1227 O ILE A 158 −4.701 −5.023 −30.081 1.00 19.04 ATOM 1228 NVAL A 159 −6.840 −5.704 −30.359 1.00 19.39 ATOM 1229 CA VAL A 159 −7.201−5.624 −28.953 1.00 19.15 ATOM 1230 CB VAL A 159 −8.687 −6.026 −28.7441.00 19.06 ATOM 1231 CG1 VAL A 159 −9.046 −6.028 −27.253 1.00 20.39 ATOM1232 CG2 VAL A 159 −9.604 −5.090 −29.511 1.00 20.08 ATOM 1233 C VAL A159 −6.280 −6.501 −28.105 1.00 19.39 ATOM 1234 O VAL A 159 −5.794 −6.089−27.036 1.00 18.63 ATOM 1235 N ARG A 160 −6.022 −7.721 −28.585 1.0018.93 ATOM 1236 CA ARG A 160 −5.171 −8.633 −27.833 1.00 19.64 ATOM 1237CB ARG A 160 −5.078 −10.005 −28.513 1.00 19.17 ATOM 1238 CG ARG A 160−4.064 −10.942 −27.872 1.00 21.41 ATOM 1239 CD ARG A 160 −3.978 −12.278−28.637 1.00 23.47 ATOM 1240 NE ARG A 160 −3.542 −12.066 −30.021 1.0029.25 ATOM 1241 CZ ARG A 160 −3.963 −12.771 −31.074 1.00 33.46 ATOM 1242NH1 ARG A 160 −4.839 −13.764 −30.929 1.00 36.48 ATOM 1243 NH2 ARG A 160−3.501 −12.489 −32.289 1.00 34.33 ATOM 1244 C ARG A 160 −3.785 −8.058−27.580 1.00 18.24 ATOM 1245 O APG A 160 −3.262 −8.233 −26.517 1.0018.13 ATOM 1246 N ASN A 161 −3.182 −7.371 −28.551 1.00 18.57 ATOM 1247CA ASN A 161 −1.875 −6.717 −28.289 1.00 18.40 ATOM 1248 CB ASN A 161−1.344 −6.052 −29.561 1.00 18.82 ATOM 1249 CG ASN A 161 −0.772 −7.055−30.549 1.00 20.68 ATOM 1250 OD1 ASN A 161 −0.240 −8.097 −30.149 1.0022.60 ATOM 1251 ND2 ASN A 161 −0.883 −6.751 −31.837 1.00 19.81 ATOM 1252C ASN A 161 −1.946 −5.656 −27.182 1.00 18.36 ATOM 1253 O ASN A 161−1.078 −5.581 −26.313 1.00 17.42 ATOM 1254 N ASP A 162 −2.982 −4.816−27.233 1.00 17.86 ATOM 1255 CA ASP A 162 −3.163 −3.782 −26.194 1.0016.85 ATOM 1256 CB ASP A 162 −4.293 −2.821 −26.586 1.00 16.71 ATOM 1257CG ASP A 162 −3.851 −1.791 −27.623 1.00 17.46 ATOM 1258 OD1 ASP A 162−2.648 −1.440 −27.681 1.00 16.98 ATOM 1259 OD2 ASP A 162 −4.719 −1.333−28.388 1.00 18.89 ATOM 1260 C ASP A 162 −3.421 −4.354 −24.799 1.0016.51 ATOM 1261 O ASP A 162 −2.846 −3.897 −23.822 1.00 15.71 ATOM 1262 NLEU A 163 −4.278 −5.371 −24.715 1.00 16.83 ATOM 1263 CA LEU A 163 −4.532−6.071 −23.459 1.00 16.55 ATOM 1264 CB LEU A 163 −5.661 −7.088 −23.6371.00 16.96 ATOM 1265 CG LEU A 163 −7.030 −6.506 −23.975 1.00 19.29 ATOM1266 CD1 LEU A 163 −8.007 −7.663 −24.227 1.00 19.71 ATOM 1267 CD2 LEU A163 −7.484 −5.631 −22.818 1.00 21.19 ATOM 1268 C LEU A 163 −3.279 −6.750−22.883 1.00 16.19 ATOM 1269 O LEU A 163 −3.035 −6.690 −21.688 1.0015.47 ATOM 1270 N ASN A 164 −2.495 −7.401 −23.748 1.00 16.85 ATOM 1271CA ASN A 164 −1.251 −8.040 −23.305 1.00 16.44 ATOM 1272 CB ASN A 164−0.602 −8.836 −24.450 1.00 17.06 ATOM 1273 CG ASN A 164 −1.333 −10.153−24.718 1.00 19.63 ATOM 1274 OD1 ASN A 164 −2.274 −10.513 −23.982 1.0020.97 ATOM 1275 ND2 ASN A 164 −0.903 −10.881 −25.756 1.00 19.79 ATOM1276 C ASN A 164 −0.301 −7.022 −22.761 1.00 16.80 ATOM 1277 O ASN A 1640.349 −7.261 −21.751 1.00 15.97 ATOM 1278 N TYR A 165 −0.250 −5.860−23.415 1.00 16.45 ATOM 1279 CA TYR A 165 0.573 −4.744 −22.930 1.0016.15 ATOM 1280 CB TYR A 165 0.420 −3.508 −23.844 1.00 16.63 ATOM 1281CG TYR A 165 1.286 −2.356 −23.391 1.00 16.41 ATOM 1282 CD1 TYR A 1650.838 −1.459 −22.404 1.00 17.94 ATOM 1283 CE1 TYR A 165 1.651 −0.402−21.958 1.00 17.84 ATOM 1284 CZ TYR A 165 2.916 −0.223 −22.517 1.0018.36 ATOM 1285 OH TYR A 165 3.699 0.841 −22.091 1.00 16.54 ATOM 1286CE2 TYR A 165 3.383 −1.105 −23.502 1.00 16.73 ATOM 1287 CD2 TYR A 1652.552 −2.157 −23.942 1.00 16.01 ATOM 1288 C TYR A 165 0.198 −4.366−21.503 1.00 16.26 ATOM 1289 O TYR A 165 1.073 −4.218 −20.650 1.00 15.43ATOM 1290 N VAL A 166 −1.104 −4.177 −21.258 1.00 16.75 ATOM 1291 CA VALA 166 −1.600 −3.786 −19.933 1.00 17.39 ATOM 1292 CB VAL A 166 −3.124−3.479 −19.986 1.00 17.57 ATOM 1293 CG1 VAL A 166 −3.712 −3.197 −18.5821.00 19.25 ATOM 1294 CG2 VAL A 166 −3.363 −2.272 −20.909 1.00 16.49 ATOM1295 C VAL A 166 −1.258 −4.829 −18.865 1.00 17.83 ATOM 1296 O VAL A 166−0.741 −4.483 −17.792 1.00 18.00 ATOM 1297 N ALA A 167 −1.520 −6.099−19.188 1.00 18.26 ATOM 1298 CA ALA A 167 −1.233 −7.218 −18.285 1.0018.92 ATOM 1299 CB ALA A 167 −1.716 −8.532 −18.899 1.00 18.25 ATOM 1300C ALA A 167 0.251 −7.325 −17.956 1.00 19.08 ATOM 1301 O ALA A 167 0.611−7.757 −16.854 1.00 20.02 ATOM 1302 N GLN A 168 1.097 −6.955 −18.9201.00 19.13 ATOM 1303 CA GLN A 168 2.558 −7.022 −18.749 1.00 19.21 ATOM1304 CB GLN A 168 3.218 −7.201 −20.115 1.00 19.08 ATOM 1305 CG GLN A 1684.739 −7.373 −20.053 1.00 20.55 ATOM 1306 CD GLN A 168 5.337 −7.891−21.355 1.00 20.26 ATOM 1307 OE1 GLN A 168 4.634 −8.378 −22.238 1.0022.69 ATOM 1308 NE2 GLN A 168 6.643 −7.772 −21.476 1.00 23.10 ATOM 1309C GLN A 168 3.182 −5.807 −18.048 1.00 19.60 ATOM 1310 O GLN A 168 4.104−5.942 −17.205 1.00 18.87 ATOM 1311 N TYR A 169 2.709 −4.609 −18.4041.00 19.23 ATOM 1312 CA TYR A 169 3.399 −3.377 −18.011 1.00 19.45 ATOM1313 CB TYR A 169 3.760 −2.560 −19.266 1.00 20.43 ATOM 1314 CG TYR A 1694.773 −3.203 −20.203 1.00 21.30 ATOM 1315 CD1 TYR A 169 6.125 −3.243−19.872 1.00 23.63 ATOM 1316 CE1 TYR A 169 7.065 −3.822 −20.723 1.0024.59 ATOM 1317 CZ TYR A 169 6.651 −4.359 −21.926 1.00 23.11 ATOM 1318OH TYR A 169 7.580 −4.924 −22.779 1.00 25.26 ATOM 1319 CE2 TYR A 1695.309 −4.330 −22.288 1.00 22.49 ATOM 1320 CD2 TYR A 169 4.375 −3.754−21.422 1.00 20.87 ATOM 1321 C TYR A 169 2.675 −2.449 −17.015 1.00 19.49ATOM 1322 O TYR A 169 3.205 −1.386 −16.691 1.00 19.69 ATOM 1323 N TRP A170 1.508 −2.850 −16.498 1.00 18.91 ATOM 1324 CA TRP A 170 0.735 −1.981−15.588 1.00 19.29 ATOM 1325 CB TRP A 170 −0.610 −2.626 −15.208 1.0018.85 ATOM 1326 CG TRP A 170 −0.489 −3.743 −14.215 1.00 21.04 ATOM 1327CD1 TRP A 170 −0.342 −5.083 −14.489 1.00 20.67 ATOM 1328 NE1 TRP A 170−0.259 −5.793 −13.317 1.00 22.09 ATOM 1329 CE2 TRP A 170 −0.336 −4.928−12.258 1.00 19.19 ATOM 1330 CD2 TRP A 170 −0.481 −3.621 −12.789 1.0020.35 ATOM 1331 CE3 TRP A 170 −0.582 −2.530 −11.905 1.00 19.61 ATOM 1332CZ3 TRP A 170 −0.546 −2.769 −10.542 1.00 22.33 ATOM 1333 CH2 TRP A 170−0.404 −4.090 −10.038 1.00 21.08 ATOM 1334 CZ2 TRP A 170 −0.297 −5.179−10.884 1.00 20.76 ATOM 1335 C TRP A 170 1.526 −1.592 −14.336 1.00 19.20ATOM 1336 O TRP A 170 1.395 −0.475 −13.808 1.00 19.24 ATOM 1337 N ASN A171 2.371 −2.504 −13.858 1.00 19.13 ATOM 1338 CA ASN A 171 3.054 −2.280−12.596 1.00 20.12 ATOM 1339 CB ASN A 171 3.178 −3.603 −11.820 1.0020.59 ATOM 1340 CG ASN A 171 3.646 −3.419 −10.392 1.00 22.31 ATOM 1341OD1 ASN A 171 4.531 −4.155 −9.938 1.00 23.68 ATOM 1342 ND2 ASN A 1713.081 −2.435 −9.684 1.00 18.77 ATOM 1343 C ASN A 171 4.392 −1.557−12.797 1.00 20.93 ATOM 1344 O ASN A 171 5.333 −1.724 −12.022 1.00 20.35ATOM 1345 N GLN A 172 4.449 −0.712 −13.826 1.00 20.64 ATOM 1346 CA GLN A172 5.644 0.061 −14.156 1.00 22.31 ATOM 1347 CB GLN A 172 6.262 −0.452−15.469 1.00 22.04 ATOM 1348 CG GLN A 172 6.784 −1.895 −15.312 1.0025.79 ATOM 1349 CD GLN A 172 7.536 −2.450 −16.515 1.00 27.61 ATOM 1350OE1 GLN A 172 8.276 −1.735 −17.214 1.00 35.65 ATOM 1351 NE2 GLN A 1727.367 −3.755 −16.752 1.00 33.80 ATOM 1352 C GLN A 172 5.287 1.539−14.268 1.00 21.36 ATOM 1353 O GLN A 172 4.175 1.867 −14.704 1.00 21.04ATOM 1354 N THR A 173 6.209 2.417 −13.871 1.00 19.73 ATOM 1355 CA THR A173 5.948 3.871 −13.928 1.00 20.07 ATOM 1356 CB THR A 173 7.001 4.703−13.168 1.00 19.48 ATOM 1357 OG1 THR A 173 8.300 4.427 −13.707 1.0021.56 ATOM 1358 CG2 THR A 173 6.988 4.375 −11.690 1.00 20.86 ATOM 1359 CTHR A 173 5.913 4.347 −15.375 1.00 18.53 ATOM 1360 O THR A 173 6.3953.665 −16.292 1.00 18.48 ATOM 1361 N GLY A 174 5.345 5.528 −15.582 1.0018.38 ATOM 1362 CA GLY A 174 5.363 6.149 −16.903 1.00 17.13 ATOM 1363 CGLY A 174 4.760 7.522 −16.736 1.00 16.77 ATOM 1364 O GLY A 174 4.4627.939 −15.605 1.00 16.87 ATOM 1365 N PHE A 175 4.571 8.223 −17.849 1.0014.68 ATOM 1366 CA PHE A 175 4.004 9.577 −17.776 1.00 14.68 ATOM 1367 CBPHE A 175 4.522 10.432 −18.948 1.00 15.16 ATOM 1368 CG PHE A 175 5.94310.847 −18.756 1.00 15.28 ATOM 1369 CD1 PHE A 175 6.981 10.000 −19.1441.00 18.04 ATOM 1370 CE1 PHE A 175 8.313 10.359 −18.915 1.00 19.53 ATOM1371 CZ PHE A 175 8.609 11.582 −18.278 1.00 19.12 ATOM 1372 CE2 PHE A175 7.571 12.429 −17.876 1.00 18.63 ATOM 1373 CD2 PHE A 175 6.247 12.054−18.113 1.00 17.67 ATOM 1374 C PHE A 175 2.483 9.584 −17.655 1.00 14.11ATOM 1375 O PHE A 175 1.799 8.683 −18.175 1.00 14.32 ATOM 1376 N ASP A176 1.972 10.591 −16.938 1.00 14.79 ATOM 1377 CA ASP A 176 0.541 10.764−16.713 1.00 14.45 ATOM 1378 CB ASP A 176 0.297 11.661 −15.506 1.0013.51 ATOM 1379 CG ASP A 176 0.685 13.126 −15.760 1.00 14.99 ATOM 1380OD1 ASP A 176 1.774 13.399 −16.329 1.00 14.32 ATOM 1381 OD2 ASP A 176−0.112 14.012 −15.376 1.00 15.08 ATOM 1382 C ASP A 176 −0.143 11.343−17.962 1.00 14.21 ATOM 1383 O ASP A 176 0.525 11.641 −18.963 1.00 14.31ATOM 1384 N LEU A 177 −1.467 11.511 −17.891 1.00 13.30 ATOM 1385 CA LEUA 177 −2.235 11.981 −19.048 1.00 13.44 ATOM 1386 CB LEU A 177 −3.75211.839 −18.832 1.00 13.71 ATOM 1387 CG LEU A 177 −4.483 12.896 −18.0121.00 14.11 ATOM 1388 CD1 LEU A 177 −5.996 12.647 −18.061 1.00 13.65 ATOM1389 CD2 LEU A 177 −4.007 12.922 −16.553 1.00 14.74 ATOM 1390 C LEU A177 −1.884 13.424 −19.452 1.00 13.51 ATOM 1391 O LEU A 177 −2.131 13.813−20.600 1.00 13.74 ATOM 1392 N TRP A 178 −1.319 14.206 −18.521 1.0012.53 ATOM 1393 CA TRP A 178 −0.804 15.553 −18.855 1.00 12.95 ATOM 1394CB TRP A 178 −0.890 16.507 −17.660 1.00 12.67 ATOM 1395 CG TRP A 178−2.247 16.549 −17.005 1.00 13.10 ATOM 1396 CD1 TRP A 178 −2.504 16.508−15.662 1.00 13.10 ATOM 1397 NE1 TRP A 178 −3.856 16.568 −15.440 1.0012.14 ATOM 1398 CE2 TRP A 178 −4.501 16.646 −16.646 1.00 13.00 ATOM 1399CD2 TRP A 178 −3.516 16.641 −17.657 1.00 12.53 ATOM 1400 CE3 TRP A 178−3.919 16.715 −19.002 1.00 12.38 ATOM 1401 CZ3 TRP A 178 −5.309 16.813−19.290 1.00 13.91 ATOM 1402 CH2 TRP A 178 −6.262 16.804 −18.257 1.0013.52 ATOM 1403 CZ2 TRP A 178 −5.883 16.718 −16.930 1.00 13.97 ATOM 1404C TRP A 178 0.632 15.565 −19.400 1.00 13.35 ATOM 1405 O TRP A 178 1.14716.641 −19.756 1.00 13.76 ATOM 1406 N GLU A 179 1.255 14.387 −19.4471.00 13.33 ATOM 1407 CA GLU A 179 2.532 14.151 −20.117 1.00 13.32 ATOM1408 CB GLU A 179 2.503 14.632 −21.582 1.00 12.64 ATOM 1409 CG GLU A 1791.165 14.344 −22.280 1.00 13.03 ATOM 1410 CD GLU A 179 1.274 14.434−23.785 1.00 14.68 ATOM 1411 OE1 GLU A 179 0.895 15.478 −24.340 1.0015.98 ATOM 1412 OE2 GLU A 179 1.730 13.457 −24.405 1.00 15.44 ATOM 1413C GLU A 179 3.667 14.853 −19.374 1.00 15.00 ATOM 1414 O GLU A 179 4.62615.292 −20.004 1.00 14.90 ATOM 1415 N GLU A 180 3.561 14.932 −18.0481.00 14.78 ATOM 1416 CA GLU A 180 4.476 15.745 −17.246 1.00 16.76 ATOM1417 CB GLU A 180 3.719 16.928 −16.630 1.00 16.95 ATOM 1418 CG GLU A 1803.282 17.972 −17.654 1.00 18.69 ATOM 1419 CD GLU A 180 2.240 18.969−17.122 1.00 19.72 ATOM 1420 OE1 GLU A 180 1.587 18.715 −16.077 1.0019.00 ATOM 1421 OE2 GLU A 180 2.076 20.020 −17.793 1.00 24.62 ATOM 1422C GLU A 180 5.124 14.954 −16.104 1.00 16.50 ATOM 1423 O GLU A 180 6.26515.202 −15.750 1.00 17.36 ATOM 1424 N VAL A 181 4.364 14.056 −15.4881.00 16.77 ATOM 1425 CA VAL A 181 4.775 13.426 −14.218 1.00 16.87 ATOM1426 CB VAL A 181 3.672 13.555 −13.130 1.00 16.78 ATOM 1427 CG1 VAL A181 4.030 12.732 −11.893 1.00 18.56 ATOM 1428 CG2 VAL A 181 3.490 15.008−12.726 1.00 17.21 ATOM 1429 C VAL A 181 5.057 11.953 −14.451 1.00 17.22ATOM 1430 O VAL A 181 4.177 11.205 −14.825 1.00 16.93 ATOM 1431 N ASN A182 6.290 11.532 −14.201 1.00 18.39 ATOM 1432 CA ASN A 182 6.674 10.123−14.394 1.00 18.57 ATOM 1433 CB ASN A 182 8.136 10.079 −14.845 1.0019.77 ATOM 1434 CG ASN A 182 8.665 8.669 −15.056 1.00 23.96 ATOM 1435OD1 ASN A 182 9.881 8.470 −15.058 1.00 33.20 ATOM 1436 ND2 ASN A 1827.794 7.706 −15.258 1.00 23.63 ATOM 1437 C ASN A 182 6.440 9.375 −13.0731.00 18.31 ATOM 1438 O ASN A 182 7.132 9.621 −12.087 1.00 18.80 ATOM1439 N GLY A 183 5.436 8.508 −13.034 1.00 16.83 ATOM 1440 CA GLY A 1835.091 7.828 −11.790 1.00 15.98 ATOM 1441 C GLY A 183 3.989 6.837 −12.0331.00 15.59 ATOM 1442 O GLY A 183 3.937 6.228 −13.117 1.00 15.30 ATOM1443 N SER A 184 3.119 6.670 −11.035 1.00 14.95 ATOM 1444 CA SER A 1841.927 5.823 −11.151 1.00 15.45 ATOM 1445 CB SER A 184 1.844 4.792−10.017 1.00 15.76 ATOM 1446 OG SER A 184 2.998 3.935 −10.027 1.00 17.33ATOM 1447 C SER A 184 0.731 6.758 −11.073 1.00 15.14 ATOM 1448 O SER A184 0.646 7.546 −10.148 1.00 15.83 ATOM 1449 N SER A 185 −0.151 6.706−12.066 1.00 14.73 ATOM 1450 CA SER A 185 −1.169 7.755 −12.190 1.0013.87 ATOM 1451 CB SER A 185 −0.991 8.535 −13.515 1.00 14.73 ATOM 1452OG SER A 185 −1.793 9.721 −13.544 1.00 14.93 ATOM 1453 C SER A 185−2.551 7.140 −12.127 1.00 13.49 ATOM 1454 O SER A 185 −2.834 6.134−12.792 1.00 13.35 ATOM 1455 N PHE A 186 −3.427 7.782 −11.354 1.00 13.72ATOM 1456 CA PHE A 186 −4.764 7.275 −11.092 1.00 13.43 ATOM 1457 CB PHEA 186 −5.511 8.319 −10.260 1.00 13.58 ATOM 1458 CG PHE A 186 −6.8077.839 −9.662 1.00 13.52 ATOM 1459 CD1 PHE A 186 −6.819 6.873 −8.655 1.0016.11 ATOM 1460 CE1 PHE A 186 −8.004 6.489 −8.036 1.00 17.80 ATOM 1461CZ PHE A 186 −9.214 7.062 −8.442 1.00 16.18 ATOM 1462 CE2 PHE A 186−9.211 8.051 −9.432 1.00 15.77 ATOM 1463 CD2 PHE A 186 −8.003 8.435−10.030 1.00 14.62 ATOM 1464 C PHE A 186 −5.552 6.946 −12.372 1.00 13.57ATOM 1465 O PHE A 186 −6.053 5.839 −12.524 1.00 13.36 ATOM 1466 N PHE A187 −5.693 7.927 −13.267 1.00 12.53 ATOM 1467 CA PHE A 187 −6.416 7.762−14.527 1.00 12.84 ATOM 1468 CB PHE A 187 −6.284 9.056 −15.356 1.0011.69 ATOM 1469 CG PHE A 187 −6.949 9.016 −16.711 1.00 13.25 ATOM 1470CD1 PHE A 187 −8.284 9.338 −16.855 1.00 12.86 ATOM 1471 CE1 PHE A 187−8.893 9.342 −18.102 1.00 14.12 ATOM 1472 CZ PHE A 187 −8.139 9.041−19.236 1.00 14.05 ATOM 1473 CE2 PHE A 187 −6.806 8.721 −19.111 1.0014.30 ATOM 1474 CD2 PHE A 187 −6.206 8.711 −17.857 1.00 15.10 ATOM 1475C PHE A 187 −5.887 6.563 −15.318 1.00 12.69 ATOM 1476 O PHE A 187 −6.6665.837 −15.932 1.00 14.00 ATOM 1477 N THR A 188 −4.571 6.357 −15.294 1.0012.97 ATOM 1478 CA THR A 188 −3.938 5.302 −16.084 1.00 13.65 ATOM 1479CB THR A 188 −2.411 5.541 −16.104 1.00 13.69 ATOM 1480 OG1 THR A 188−2.158 6.789 −16.753 1.00 15.37 ATOM 1481 CG2 THR A 188 −1.648 4.432−16.833 1.00 13.24 ATOM 1482 C THR A 188 −4.284 3.929 −15.478 1.00 14.12ATOM 1483 O THR A 188 −4.766 3.039 −16.173 1.00 14.40 ATOM 1484 N VAL A189 −4.066 3.798 −14.173 1.00 13.34 ATOM 1485 CA VAL A 189 −4.348 2.543−13.446 1.00 14.76 ATOM 1486 CB VAL A 189 −3.893 2.612 −11.958 1.0014.90 ATOM 1487 CG1 VAL A 189 −4.331 1.334 −11.186 1.00 16.95 ATOM 1488CG2 VAL A 189 −2.374 2.799 −11.865 1.00 15.31 ATOM 1489 C VAL A 189−5.836 2.167 −13.560 1.00 14.48 ATOM 1490 O VAL A 189 −6.159 1.024−13.853 1.00 14.65 ATOM 1491 N ALA A 190 −6.732 3.146 −13.372 1.00 13.77ATOM 1492 CA ALA A 190 −8.171 2.858 −13.351 1.00 13.46 ATOM 1493 CB ALAA 190 −8.996 4.128 −12.922 1.00 12.74 ATOM 1494 C ALA A 190 −8.614 2.388−14.706 1.00 13.27 ATOM 1495 O ALA A 190 −9.432 1.479 −14.815 1.00 13.24ATOM 1496 N ASN A 191 −8.093 3.017 −15.760 1.00 12.50 ATOM 1497 CA ASN A191 −8.438 2.598 −17.127 1.00 13.01 ATOM 1498 CB ASN A 191 −8.122 3.707−18.137 1.00 12.65 ATOM 1499 CG ASN A 191 −9.191 4.781 −18.118 1.0014.08 ATOM 1500 OD1 ASN A 191 −10.319 4.541 −18.554 1.00 16.39 ATOM 1501ND2 ASN A 191 −8.857 5.955 −17.583 1.00 17.21 ATOM 1502 C ASN A 191−7.815 1.259 −17.521 1.00 13.93 ATOM 1503 O ASN A 191 −8.412 0.490−18.270 1.00 13.87 ATOM 1504 N GLN A 192 −6.636 0.995 −16.980 1.00 14.06ATOM 1505 CA GLN A 192 −5.988 −0.311 −17.139 1.00 14.96 ATOM 1506 CB GLNA 192 −4.575 −0.274 −16.552 1.00 14.33 ATOM 1507 CG GLN A 192 −3.5550.435 −17.500 1.00 13.64 ATOM 1508 CD GLN A 192 −2.206 0.635 −16.8571.00 15.33 ATOM 1509 OE1 GLN A 192 −2.074 0.568 −15.646 1.00 15.48 ATOM1510 NE2 GLN A 192 −1.182 0.925 −17.682 1.00 16.10 ATOM 1511 C GLN A 192−6.855 −1.411 −16.519 1.00 15.11 ATOM 1512 O GLN A 192 −7.076 −2.457−17.141 1.00 16.04 ATOM 1513 N HIS A 193 −7.398 −1.140 −15.329 1.0015.81 ATOM 1514 CA HIS A 193 −8.314 −2.069 −14.668 1.00 16.01 ATOM 1515CB HIS A 193 −8.746 −1.586 −13.281 1.00 16.72 ATOM 1516 CG HIS A 193−9.806 −2.454 −12.669 1.00 17.39 ATOM 1517 ND1 HIS A 193 −11.113 −2.039−12.505 1.00 18.05 ATOM 1518 CE1 HIS A 193 −11.821 −3.028 −11.983 1.0018.70 ATOM 1519 NE2 HIS A 193 −11.023 −4.071 −11.814 1.00 17.20 ATOM1520 CD2 HIS A 193 −9.758 −3.739 −12.242 1.00 18.04 ATOM 1521 C HIS A193 −9.536 −2.343 −15.521 1.00 15.85 ATOM 1522 O HIS A 193 −9.898 −3.501−15.732 1.00 15.70 ATOM 1523 N ARG A 194 −10.185 −1.285 −15.995 1.0015.27 ATOM 1524 CA ARG A 194 −11.349 −1.437 −16.852 1.00 15.29 ATOM 1525CB ARG A 194 −11.922 −0.073 −17.234 1.00 14.30 ATOM 1526 CG ARG A 194−13.029 −0.212 −18.239 1.00 14.46 ATOM 1527 CD ARG A 194 −13.614 1.102−18.723 1.00 15.43 ATOM 1528 NE ARG A 194 −14.589 0.780 −19.767 1.0015.58 ATOM 1529 CZ ARG A 194 −15.624 1.539 −20.125 1.00 17.92 ATOM 1530NH1 ARG A 194 −15.815 2.744 −19.576 1.00 14.60 ATOM 1531 NH2 ARG A 194−16.451 1.095 −21.060 1.00 16.15 ATOM 1532 C ARG A 194 −11.047 −2.258−18.111 1.00 15.74 ATOM 1533 O ARG A 194 −11.842 −3.120 −18.504 1.0015.56 ATOM 1534 N ALA A 195 −9.918 −1.967 −18.758 1.00 15.60 ATOM 1535CA ALA A 195 −9.562 −2.638 −20.004 1.00 15.90 ATOM 1536 CB ALA A 195−8.254 −2.042 −20.591 1.00 15.40 ATOM 1537 C ALA A 195 −9.436 −4.150−19.798 1.00 15.65 ATOM 1538 O ALA A 195 −9.959 −4.929 −20.610 1.0016.79 ATOM 1539 N LEU A 196 −8.763 −4.550 −18.721 1.00 16.36 ATOM 1540CA LEU A 196 −8.552 −5.976 −18.423 1.00 17.02 ATOM 1541 CB LEU A 196−7.625 −6.126 −17.235 1.00 16.96 ATOM 1542 CG LEU A 196 −6.167 −5.744−17.532 1.00 16.96 ATOM 1543 CD1 LEU A 196 −5.375 −5.857 −16.252 1.0018.93 ATOM 1544 CD2 LEU A 196 −5.590 −6.636 −18.630 1.00 20.38 ATOM 1545C LEU A 196 −9.877 −6.685 −18.167 1.00 17.92 ATOM 1546 O LEU A 196−10.102 −7.795 −18.643 1.00 18.98 ATOM 1547 N VAL A 197 −10.779 −6.014−17.454 1.00 18.51 ATOM 1548 CA VAL A 197 −12.112 −6.560 −17.181 1.0018.81 ATOM 1549 CB VAL A 197 −12.875 −5.702 −16.130 1.00 18.26 ATOM 1550CG1 VAL A 197 −14.340 −6.173 −15.994 1.00 21.18 ATOM 1551 CG2 VAL A 197−12.149 −5.784 −14.778 1.00 19.79 ATOM 1552 C VAL A 197 −12.924 −6.779−18.462 1.00 19.05 ATOM 1553 O VAL A 197 −13.456 −7.884 −18.693 1.0018.62 ATOM 1554 N GLU A 198 −13.010 −5.752 −19.308 1.00 18.43 ATOM 1555CA GLU A 198 −13.747 −5.873 −20.556 1.00 19.38 ATOM 1556 CB GLU A 198−13.849 −4.517 −21.241 1.00 19.38 ATOM 1557 CG GLU A 198 −14.609 −3.530−20.417 1.00 20.22 ATOM 1558 CD GLU A 198 −15.334 −2.537 −21.298 1.0022.66 ATOM 1559 OE1 GLU A 198 −16.313 −2.940 −21.940 1.00 22.16 ATOM1560 OE2 GLU A 198 −14.924 −1.369 −21.342 1.00 22.92 ATOM 1561 C GLU A198 −13.094 −6.861 −21.509 1.00 19.78 ATOM 1562 O GLU A 198 −13.780−7.506 −22.303 1.00 20.29 ATOM 1563 N GLY A 199 −11.770 −6.944 −21.4351.00 19.78 ATOM 1564 CA GLY A 199 −10.998 −7.823 −22.314 1.00 20.88 ATOM1565 C GLY A 199 −11.288 −9.285 −21.986 1.00 21.53 ATOM 1566 O GLY A 199−11.546 −10.083 −22.879 1.00 22.36 ATOM 1567 N ALA A 200 −11.256 −9.615−20.702 1.00 21.79 ATOM 1568 CA ALA A 200 −11.605 −10.956 −20.234 1.0022.44 ATOM 1569 CB ALA A 200 −11.463 −11.038 −18.728 1.00 22.21 ATOM1570 C ALA A 200 −13.016 −11.329 −20.696 1.00 22.54 ATOM 1571 O ALA A200 −13.237 −12.419 −21.214 1.00 22.25 ATOM 1572 N THR A 201 −13.965−10.403 −20.573 1.00 22.56 ATOM 1573 CA THR A 201 −15.345 −10.671−20.989 1.00 22.77 ATOM 1574 CB THR A 201 −16.302 −9.527 −20.551 1.0022.83 ATOM 1575 OG1 THR A 201 −16.219 −9.387 −19.134 1.00 24.92 ATOM1576 CG2 THR A 201 −17.756 −9.819 −20.929 1.00 23.76 ATOM 1577 C THR A201 −15.435 −10.905 −22.485 1.00 22.78 ATOM 1578 O THR A 201 −16.099−11.851 −22.925 1.00 22.95 ATOM 1579 N LEU A 202 −14.760 −10.069 −23.2751.00 21.78 ATOM 1580 CA LEU A 202 −14.805 −10.236 −24.717 1.00 22.62ATOM 1581 CB LEU A 202 −14.149 −9.055 −25.434 1.00 22.14 ATOM 1582 CGLEU A 202 −14.142 −9.107 −26.964 1.00 23.10 ATOM 1583 CD1 LEU A 202−15.544 −9.198 −27.564 1.00 24.20 ATOM 1584 CD2 LEU A 202 −13.346 −7.938−27.570 1.00 22.53 ATOM 1585 C LEU A 202 −14.139 −11.552 −25.151 1.0023.15 ATOM 1586 O LEU A 202 −14.649 −12.245 −26.036 1.00 22.90 ATOM 1587N ALA A 203 −13.019 −11.883 −24.510 1.00 23.38 ATOM 1588 CA ALA A 203−12.300 −13.129 −24.787 1.00 24.07 ATOM 1589 CB ALA A 203 −11.076−13.229 −23.913 1.00 23.57 ATOM 1590 C ALA A 203 −13.211 −14.354 −24.5691.00 24.38 ATOM 1591 O ALA A 203 −13.264 −15.244 −25.411 1.00 25.21 ATOM1592 N ALA A 204 −13.920 −14.363 −23.447 1.00 25.20 ATOM 1593 CA ALA A204 −14.849 −15.442 −23.093 1.00 26.63 ATOM 1594 CB ALA A 204 −15.450−15.186 −21.727 1.00 26.12 ATOM 1595 C ALA A 204 −15.939 −15.583 −24.1501.00 27.48 ATOM 1596 O ALA A 204 −16.267 −16.687 −24.564 1.00 28.39 ATOM1597 N THR A 205 −16.494 −14.461 −24.593 1.00 27.71 ATOM 1598 CA THR A205 −17.497 −14.470 −25.652 1.00 28.39 ATOM 1599 CB THR A 205 −18.088−13.051 −25.855 1.00 28.42 ATOM 1600 OG1 THR A 205 −18.669 −12.631−24.622 1.00 29.32 ATOM 1601 CG2 THR A 205 −19.150 −13.051 −26.932 1.0027.32 ATOM 1602 C THR A 205 −16.968 −15.004 −26.981 1.00 28.72 ATOM 1603O THR A 205 −17.697 −15.690 −27.719 1.00 29.06 ATOM 1604 N LEU A 206−15.712 −14.698 −27.288 1.00 28.58 ATOM 1605 CA LEU A 206 −15.122−15.122 −28.539 1.00 29.40 ATOM 1606 CB LEU A 206 −14.034 −14.144−29.001 1.00 29.40 ATOM 1607 CG LEU A 206 −14.438 −12.694 −29.322 1.0029.97 ATOM 1608 CD1 LEU A 206 −13.212 −11.899 −29.755 1.00 30.30 ATOM1609 CD2 LEU A 206 −15.561 −12.629 −30.375 1.00 29.58 ATOM 1610 C LEU A206 −14.540 −16.538 −28.489 1.00 29.55 ATOM 1611 O LEU A 206 −14.118−17.054 −29.521 1.00 30.16 ATOM 1612 N GLY A 207 −14.500 −17.145 −27.3071.00 30.30 ATOM 1613 CA GLY A 207 −13.786 −18.419 −27.122 1.00 30.91ATOM 1614 C GLY A 207 −12.294 −18.274 −27.375 1.00 31.55 ATOM 1615 O GLYA 207 −11.654 −19.173 −27.935 1.00 31.31 ATOM 1616 N GLN A 208 −11.746−17.115 −26.989 1.00 31.08 ATOM 1617 CA GLN A 208 −10.311 −16.877−27.031 1.00 31.10 ATOM 1618 CB GLN A 208 −9.999 −15.540 −27.703 1.0031.08 ATOM 1619 CG GLN A 208 −10.451 −15.455 −29.142 1.00 33.86 ATOM1620 CD GLN A 208 −9.469 −16.059 −30.126 1.00 38.19 ATOM 1621 OE1 GLN A208 −9.686 −15.999 −31.335 1.00 41.96 ATOM 1622 NE2 GLN A 208 −8.386−16.633 −29.626 1.00 38.96 ATOM 1623 C GLN A 208 −9.765 −16.909 −25.6111.00 30.45 ATOM 1624 O GLN A 208 −10.516 −17.048 −24.658 1.00 30.63 ATOM1625 N SER A 209 −8.451 −16.816 −25.469 1.00 29.96 ATOM 1626 CA SER A209 −7.841 −16.898 −24.160 1.00 30.04 ATOM 1627 CB SER A 209 −6.382−17.343 −24.297 1.00 30.04 ATOM 1628 OG SER A 209 −5.763 −17.371 −23.0301.00 32.75 ATOM 1629 C SER A 209 −7.948 −15.564 −23.409 1.00 29.53 ATOM1630 O SER A 209 −7.493 −14.532 −23.908 1.00 29.85 ATOM 1631 N GLY A 210−8.545 −15.594 −22.216 1.00 28.41 ATOM 1632 CA GLY A 210 −8.745 −14.388−21.401 1.00 27.16 ATOM 1633 C GLY A 210 −8.344 −14.480 −19.938 1.0026.83 ATOM 1634 O GLY A 210 −8.425 −13.498 −19.203 1.00 26.61 ATOM 1635N SER A 211 −7.888 −15.648 −19.497 1.00 25.85 ATOM 1636 CA SER A 211−7.651 −15.867 −18.067 1.00 25.26 ATOM 1637 CB SER A 211 −7.401 −17.353−17.783 1.00 25.87 ATOM 1638 OG SER A 211 −6.315 −17.789 −18.573 1.0026.62 ATOM 1639 C SER A 211 −6.509 −15.026 −17.498 1.00 24.55 ATOM 1640O SER A 211 −6.542 −14.676 −16.311 1.00 24.46 ATOM 1641 N ALA A 212−5.505 −14.712 −18.323 1.00 23.56 ATOM 1642 CA ALA A 212 −4.423 −13.816−17.906 1.00 23.42 ATOM 1643 CB ALA A 212 −3.417 −13.622 −19.031 1.0023.66 ATOM 1644 C ALA A 212 −4.999 −12.450 −17.496 1.00 23.54 ATOM 1645O ALA A 212 −4.566 −11.848 −16.513 1.00 24.00 ATOM 1646 N TYR A 213−5.970 −11.979 −18.271 1.00 22.79 ATOM 1647 CA TYR A 213 −6.594 −10.676−18.017 1.00 22.18 ATOM 1648 CB TYR A 213 −7.453 −10.241 −19.193 1.0021.74 ATOM 1649 CG TYR A 213 −6.761 −10.345 −20.515 1.00 20.05 ATOM 1650CD1 TYR A 213 −7.461 −10.761 −21.637 1.00 20.58 ATOM 1651 CE1 TYR A 213−6.854 −10.854 −22.868 1.00 21.95 ATOM 1652 CZ TYR A 213 −5.503 −10.545−22.988 1.00 20.62 ATOM 1653 OH TYR A 213 −4.930 −10.668 −24.220 1.0021.72 ATOM 1654 CE2 TYR A 213 −4.758 −10.149 −21.888 1.00 19.76 ATOM1655 CD2 TYR A 213 −5.400 −10.038 −20.647 1.00 20.61 ATOM 1656 C TYR A213 −7.423 −10.710 −16.758 1.00 23.06 ATOM 1657 O TYR A 213 −7.320−9.804 −15.939 1.00 22.56 ATOM 1658 N SER A 214 −8.226 −11.767 −16.5781.00 23.15 ATOM 1659 CA SER A 214 −9.064 −11.832 −15.392 1.00 23.90 ATOM1660 CB SER A 214 −10.244 −12.798 −15.580 1.00 24.54 ATOM 1661 OG SER A214 −9.776 −14.085 −15.939 1.00 27.95 ATOM 1662 C SER A 214 −8.259−12.122 −14.122 1.00 23.64 ATOM 1663 O SER A 214 −8.676 −11.762 −13.0261.00 23.43 ATOM 1664 N SER A 215 −7.095 −12.743 −14.248 1.00 23.82 ATOM1665 CA SER A 215 −6.295 −12.970 −13.050 1.00 24.66 ATOM 1666 CB SER A215 −5.390 −14.205 −13.200 1.00 25.70 ATOM 1667 OG SER A 215 −4.267−13.914 −14.004 1.00 29.15 ATOM 1668 C SER A 215 −5.491 −11.739 −12.6101.00 23.98 ATOM 1669 O SER A 215 −5.217 −11.561 −11.421 1.00 24.09 ATOM1670 N VAL A 216 −5.115 −10.894 −13.566 1.00 22.89 ATOM 1671 CA VAL A216 −4.347 −9.679 −13.272 1.00 22.50 ATOM 1672 CB VAL A 216 −3.442−9.296 −14.493 1.00 22.52 ATOM 1673 CG1 VAL A 216 −2.855 −7.888 −14.3691.00 24.11 ATOM 1674 CG2 VAL A 216 −2.296 −10.317 −14.652 1.00 22.49ATOM 1675 C VAL A 216 −5.256 −8.520 −12.801 1.00 21.88 ATOM 1676 O VAL A216 −4.869 −7.745 −11.936 1.00 21.84 ATOM 1677 N ALA A 217 −6.475 −8.440−13.332 1.00 21.86 ATOM 1678 CA ALA A 217 −7.374 −7.303 −13.050 1.0021.59 ATOM 1679 CB ALA A 217 −8.721 −7.479 −13.760 1.00 21.26 ATOM 1680C ALA A 217 −7.571 −6.968 −11.558 1.00 21.55 ATOM 1681 O ALA A 217−7.447 −5.804 −11.165 1.00 21.20 ATOM 1682 N PRO A 218 −7.842 −7.988−10.701 1.00 21.95 ATOM 1683 CA PRO A 218 −8.030 −7.700 −9.282 1.0021.59 ATOM 1684 CB PRO A 218 −8.283 −9.104 −8.670 1.00 22.29 ATOM 1685CG PRO A 218 −8.789 −9.905 −9.789 1.00 22.61 ATOM 1686 CD PRO A 218−7.966 −9.435 −10.963 1.00 22.11 ATOM 1687 C PRO A 218 −6.798 −7.065−8.634 1.00 21.27 ATOM 1688 O PRO A 218 −6.928 −6.299 −7.680 1.00 20.92ATOM 1689 N GLN A 219 −5.608 −7.386 −9.141 1.00 21.17 ATOM 1690 CA GLN A219 −4.378 −6.786 −8.609 1.00 21.51 ATOM 1691 CB GLN A 219 −3.149 −7.569−9.084 1.00 22.72 ATOM 1692 CG GLN A 219 −3.113 −8.985 −8.516 1.00 24.90ATOM 1693 CD GLN A 219 −3.323 −8.982 −7.015 1.00 29.57 ATOM 1694 OE1 GLNA 219 −2.715 −8.188 −6.288 1.00 31.58 ATOM 1695 NE2 GLN A 219 −4.207−9.843 −6.545 1.00 33.22 ATOM 1696 C GLN A 219 −4.240 −5.301 −8.996 1.0021.04 ATOM 1697 O GLN A 219 −3.687 −4.490 −8.229 1.00 21.07 ATOM 1698 NVAL A 220 −4.728 −4.973 −10.187 1.00 20.00 ATOM 1699 CA VAL A 220 −4.746−3.577 −10.630 1.00 19.34 ATOM 1700 CB VAL A 220 −5.098 −3.456 −12.1281.00 19.63 ATOM 1701 CG1 VAL A 220 −4.991 −2.000 −12.581 1.00 19.15 ATOM1702 CG2 VAL A 220 −4.162 −4.342 −12.974 1.00 17.97 ATOM 1703 C VAL A220 −5.730 −2.809 −9.737 1.00 19.65 ATOM 1704 O VAL A 220 −5.419 −1.728−9.257 1.00 18.97 ATOM 1705 N LEU A 221 −6.903 −3.391 −9.490 1.00 20.12ATOM 1706 CA LEU A 221 −7.895 −2.776 −8.620 1.00 20.83 ATOM 1707 CB LEUA 221 −9.180 −3.602 −8.599 1.00 20.48 ATOM 1708 CG LEU A 221 −10.336−2.991 −7.790 1.00 22.48 ATOM 1709 CD1 LEU A 221 −10.857 −1.726 −8.4581.00 22.33 ATOM 1710 CD2 LEU A 221 −11.430 −4.011 −7.637 1.00 22.51 ATOM1711 C LEU A 221 −7.360 −2.591 −7.192 1.00 21.44 ATOM 1712 O LEU A 221−7.617 −1.578 −6.539 1.00 20.45 ATOM 1713 N CYS A 222 −6.600 −3.572−6.718 1.00 22.60 ATOM 1714 CA CYS A 222 −5.957 −3.477 −5.415 1.00 22.10ATOM 1715 CB CYS A 222 −5.159 −4.749 −5.125 1.00 23.41 ATOM 1716 SG CYSA 222 −4.975 −5.000 −3.356 1.00 28.49 ATOM 1717 C CYS A 222 −5.035−2.270 −5.317 1.00 21.22 ATOM 1718 O CYS A 222 −5.060 −1.531 −4.331 1.0021.24 ATOM 1719 N PHE A 223 −4.210 −2.070 −6.347 1.00 20.11 ATOM 1720 CAPHE A 223 −3.287 −0.955 −6.368 1.00 19.03 ATOM 1721 CB PHE A 223 −2.334−1.108 −7.558 1.00 19.15 ATOM 1722 CG PHE A 223 −1.297 −0.011 −7.6691.00 19.23 ATOM 1723 CD1 PHE A 223 −0.576 0.410 −6.558 1.00 19.90 ATOM1724 CE1 PHE A 223 0.380 1.417 −6.661 1.00 20.91 ATOM 1725 CZ PHE A 2230.645 2.017 −7.902 1.00 21.07 ATOM 1726 CE2 PHE A 223 −0.061 1.598−9.024 1.00 18.81 ATOM 1727 CD2 PHE A 223 −1.022 0.581 −8.909 1.00 18.23ATOM 1728 C PHE A 223 −4.032 0.397 −6.423 1.00 18.38 ATOM 1729 O PHE A223 −3.597 1.376 −5.818 1.00 18.27 ATOM 1730 N LEU A 224 −5.148 0.428−7.142 1.00 18.45 ATOM 1731 CA LEU A 224 −5.957 1.665 −7.277 1.00 18.42ATOM 1732 CB LEU A 224 −7.208 1.403 −8.127 1.00 17.70 ATOM 1733 CG LEU A224 −7.990 2.645 −8.610 1.00 19.73 ATOM 1734 CD1 LEU A 224 −7.133 3.427−9.584 1.00 20.37 ATOM 1735 CD2 LEU A 224 −9.302 2.228 −9.264 1.00 18.64ATOM 1736 C LEU A 224 −6.385 2.226 −5.917 1.00 18.87 ATOM 1737 O LEU A224 −6.553 3.438 −5.757 1.00 18.45 ATOM 1738 N GLN A 225 −6.578 1.336−4.944 1.00 19.17 ATOM 1739 CA GLN A 225 −6.984 1.743 −3.585 1.00 20.00ATOM 1740 CB GLN A 225 −7.340 0.511 −2.725 1.00 20.26 ATOM 1741 CG GLN A225 −8.295 −0.463 −3.409 1.00 21.22 ATOM 1742 CD GLN A 225 −9.519 0.225−3.993 1.00 22.53 ATOM 1743 OE1 GLN A 225 −10.280 0.870 −3.262 1.0023.09 ATOM 1744 NE2 GLN A 225 −9.718 0.092 −5.302 1.00 19.33 ATOM 1745 CGLN A 225 −5.944 2.599 −2.871 1.00 20.19 ATOM 1746 O GLN A 225 −6.2993.399 −2.009 1.00 20.64 ATOM 1747 N ARG A 226 −4.678 2.450 −3.253 1.0020.51 ATOM 1748 CA ARG A 226 −3.564 3.144 −2.608 1.00 21.40 ATOM 1749 CBARG A 226 −2.219 2.505 −2.990 1.00 22.72 ATOM 1750 CG ARG A 226 −2.0811.010 −2.683 1.00 26.14 ATOM 1751 CD ARG A 226 −1.806 0.741 −1.204 1.0032.16 ATOM 1752 NE ARG A 226 −3.035 0.843 −0.432 1.00 37.77 ATOM 1753 CZARG A 226 −3.997 −0.079 −0.413 1.00 41.09 ATOM 1754 NH1 ARG A 226 −5.0930.120 0.322 1.00 42.17 ATOM 1755 NH2 ARG A 226 −3.874 −1.196 −1.127 1.0042.78 ATOM 1756 C ARG A 226 −3.499 4.645 −2.915 1.00 21.23 ATOM 1757 OARG A 226 −2.723 5.358 −2.288 1.00 20.95 ATOM 1758 N PHE A 227 −4.2985.123 −3.869 1.00 20.28 ATOM 1759 CA PHE A 227 −4.280 6.545 −4.250 1.0019.67 ATOM 1760 CB PHE A 227 −4.777 6.704 −5.693 1.00 19.40 ATOM 1761 CGPHE A 227 −3.814 6.195 −6.744 1.00 18.28 ATOM 1762 CD1 PHE A 227 −3.7334.831 −7.040 1.00 18.14 ATOM 1763 CE1 PHE A 227 −2.855 4.355 −8.046 1.0018.24 ATOM 1764 CZ PHE A 227 −2.034 5.264 −8.748 1.00 16.75 ATOM 1765CE2 PHE A 227 −2.113 6.641 −8.456 1.00 18.79 ATOM 1766 CD2 PHE A 227−3.005 7.091 −7.452 1.00 17.51 ATOM 1767 C PHE A 227 −5.126 7.435 −3.3431.00 20.55 ATOM 1768 O PHE A 227 −4.967 8.659 −3.334 1.00 20.38 ATOM1769 N TRP A 228 −6.032 6.820 −2.583 1.00 20.72 ATOM 1770 CA TRP A 228−6.924 7.545 −1.671 1.00 20.71 ATOM 1771 CB TRP A 228 −8.036 6.596−1.211 1.00 20.41 ATOM 1772 CG TRP A 228 −9.030 7.228 −0.283 1.00 20.59ATOM 1773 CD1 TRP A 228 −9.243 6.915 1.040 1.00 21.81 ATOM 1774 NE1 TRPA 228 −10.255 7.722 1.557 1.00 22.69 ATOM 1775 CE2 TRP A 228 −10.7128.553 0.565 1.00 20.71 ATOM 1776 CD2 TRP A 228 −9.958 8.280 −0.607 1.0018.88 ATOM 1777 CE3 TRP A 228 −10.225 9.014 −1.772 1.00 18.79 ATOM 1778CZ3 TRP A 228 −11.209 9.986 −1.734 1.00 20.13 ATOM 1779 CH2 TRP A 228−11.937 10.242 −0.552 1.00 21.18 ATOM 1780 CZ2 TRP A 228 −11.710 9.5370.601 1.00 21.65 ATOM 1781 C TRP A 228 −6.193 8.120 −0.463 1.00 21.38ATOM 1782 O TRP A 228 −5.479 7.394 0.236 1.00 21.50 ATOM 1783 N VAL A229 −6.379 9.416 −0.209 1.00 21.95 ATOM 1784 CA VAL A 229 −5.844 10.0650.983 1.00 22.99 ATOM 1785 CB VAL A 229 −5.205 11.436 0.654 1.00 22.81ATOM 1786 CG1 VAL A 229 −4.490 12.026 1.871 1.00 23.48 ATOM 1787 CG2 VALA 229 −4.226 11.292 −0.493 1.00 23.28 ATOM 1788 C VAL A 229 −6.98410.206 2.000 1.00 24.08 ATOM 1789 O VAL A 229 −7.803 11.119 1.899 1.0023.70 ATOM 1790 N SER A 230 −7.044 9.298 2.974 1.00 25.37 ATOM 1791 CASER A 230 −8.193 9.290 3.905 1.00 27.59 ATOM 1792 CB SER A 230 −8.2548.000 4.728 1.00 27.67 ATOM 1793 OG SER A 230 −7.029 7.805 5.402 1.0031.30 ATOM 1794 C SER A 230 −8.241 10.513 4.820 1.00 27.93 ATOM 1795 OSER A 230 −9.321 10.983 5.174 1.00 28.91 ATOM 1796 N SER A 231 −7.08811.059 5.165 1.00 28.76 ATOM 1797 CA SER A 231 −7.059 12.237 6.030 1.0029.72 ATOM 1798 CB SER A 231 −5.671 12.461 6.639 1.00 30.39 ATOM 1799 OGSER A 231 −4.703 12.713 5.635 1.00 34.43 ATOM 1800 C SER A 231 −7.56613.491 5.323 1.00 29.39 ATOM 1801 O SER A 231 −8.154 14.364 5.966 1.0030.97 ATOM 1802 N GLY A 232 −7.373 13.579 4.005 1.00 27.59 ATOM 1803 CAGLY A 232 −7.867 14.728 3.247 1.00 25.22 ATOM 1804 C GLY A 232 −9.18114.518 2.493 1.00 23.25 ATOM 1805 O GLY A 232 −9.810 15.487 2.077 1.0023.19 ATOM 1806 N GLY A 233 −9.589 13.265 2.320 1.00 20.97 ATOM 1807 CAGLY A 233 −10.809 12.937 1.578 1.00 19.35 ATOM 1808 C GLY A 233 −10.67313.226 0.094 1.00 18.83 ATOM 1809 O GLY A 233 −11.636 13.655 −0.561 1.0019.20 ATOM 1810 N TYR A 234 −9.487 12.977 −0.463 1.00 17.56 ATOM 1811 CATYR A 234 −9.309 13.155 −1.915 1.00 17.17 ATOM 1812 CB TYR A 234 −8.85114.584 −2.232 1.00 18.33 ATOM 1813 CG TYR A 234 −7.441 14.876 −1.7581.00 20.39 ATOM 1814 CD1 TYR A 234 −7.203 15.340 −0.454 1.00 20.72 ATOM1815 CE1 TYR A 234 −5.905 15.594 −0.018 1.00 24.11 ATOM 1816 CZ TYR A234 −4.840 15.399 −0.897 1.00 23.78 ATOM 1817 OH TYR A 234 −3.556 15.663−0.483 1.00 26.50 ATOM 1818 CE2 TYR A 234 −5.055 14.956 −2.187 1.0024.07 ATOM 1819 CD2 TYR A 234 −6.353 14.699 −2.611 1.00 20.58 ATOM 1820C TYR A 234 −8.318 12.141 −2.482 1.00 16.60 ATOM 1821 O TYR A 234 −7.61511.465 −1.735 1.00 16.29 ATOM 1822 N VAL A 235 −8.260 12.059 −3.805 1.0015.36 ATOM 1823 CA VAL A 235 −7.325 11.164 −4.472 1.00 15.62 ATOM 1824CB VAL A 235 −7.948 10.638 −5.798 1.00 15.96 ATOM 1825 CG1 VAL A 235−6.889 9.893 −6.645 1.00 17.31 ATOM 1826 CG2 VAL A 235 −9.134 9.723−5.506 1.00 15.87 ATOM 1827 C VAL A 235 −6.011 11.904 −4.742 1.00 15.54ATOM 1828 O VAL A 235 −6.006 12.998 −5.320 1.00 15.39 ATOM 1829 N ASP A236 −4.886 11.316 −4.325 1.00 15.24 ATOM 1830 CA ASP A 236 −3.580 11.837−4.705 1.00 15.22 ATOM 1831 CB ASP A 236 −2.533 11.431 −3.652 1.00 16.45ATOM 1832 CG ASP A 236 −1.145 11.922 −3.970 1.00 18.62 ATOM 1833 OD1 ASPA 236 −0.937 12.617 −4.992 1.00 17.17 ATOM 1834 OD2 ASP A 236 −0.22311.568 −3.182 1.00 22.79 ATOM 1835 C ASP A 236 −3.303 11.256 −6.098 1.0015.06 ATOM 1836 O ASP A 236 −3.088 10.040 −6.261 1.00 15.39 ATOM 1837 NSER A 237 −3.384 12.104 −7.125 1.00 14.24 ATOM 1838 CA SER A 237 −3.51811.587 −8.503 1.00 14.09 ATOM 1839 CB SER A 237 −4.000 12.697 −9.4461.00 13.76 ATOM 1840 OG SER A 237 −5.312 13.094 −9.070 1.00 14.52 ATOM1841 C SER A 237 −2.277 10.883 −9.053 1.00 14.22 ATOM 1842 O SER A 237−2.376 10.067 −9.965 1.00 13.80 ATOM 1843 N ASN A 238 −1.099 11.219−8.521 1.00 14.70 ATOM 1844 CA ASN A 238 0.116 10.547 −8.952 1.00 15.28ATOM 1845 CB ASN A 238 0.968 11.439 −9.856 1.00 14.84 ATOM 1846 CG ASN A238 0.277 11.742 −11.176 1.00 17.08 ATOM 1847 OD1 ASN A 238 0.244 10.901−12.072 1.00 16.61 ATOM 1848 ND2 ASN A 238 −0.308 12.932 −11.278 1.0016.63 ATOM 1849 C ASN A 238 0.912 10.150 −7.736 1.00 16.07 ATOM 1850 OASN A 238 1.169 10.988 −6.890 1.00 15.88 ATOM 1851 N ILE A 239 1.2808.875 −7.659 1.00 16.09 ATOM 1852 CA ILE A 239 2.125 8.410 −6.567 1.0018.07 ATOM 1853 CB ILE A 239 1.340 7.452 −5.600 1.00 17.66 ATOM 1854 CG1ILE A 239 0.893 6.180 −6.336 1.00 18.85 ATOM 1855 CD1 ILE A 239 0.1845.109 −5.437 1.00 19.02 ATOM 1856 CG2 ILE A 239 0.116 8.194 −4.974 1.0016.96 ATOM 1857 C ILE A 239 3.381 7.760 −7.169 1.00 19.32 ATOM 1858 OILE A 239 3.571 7.797 −8.392 1.00 19.19 ATOM 1859 N ASN A 240 4.2427.170 −6.329 1.00 20.56 ATOM 1860 CA ASN A 240 5.517 6.617 −6.823 1.0022.24 ATOM 1861 CB ASN A 240 5.275 5.385 −7.717 1.00 21.93 ATOM 1862 CGASN A 240 4.874 4.153 −6.926 1.00 24.19 ATOM 1863 OD1 ASN A 240 5.2693.995 −5.772 1.00 25.98 ATOM 1864 ND2 ASN A 240 4.083 3.278 −7.538 1.0022.26 ATOM 1865 C ASN A 240 6.334 7.677 −7.571 1.00 23.27 ATOM 1866 OASN A 240 7.000 7.381 −8.562 1.00 23.15 ATOM 1867 N THR A 241 6.2618.919 −7.096 1.00 25.02 ATOM 1868 CA THR A 241 6.939 10.038 −7.729 1.0028.12 ATOM 1869 CB THR A 241 6.044 10.720 −8.817 1.00 28.09 ATOM 1870OG1 THR A 241 6.741 11.836 −9.369 1.00 28.75 ATOM 1871 CG2 THR A 2414.727 11.208 −8.231 1.00 28.30 ATOM 1872 C THR A 241 7.302 11.065 −6.6741.00 29.96 ATOM 1873 O THR A 241 6.749 11.037 −5.589 1.00 30.58 ATOM1874 N ASN A 242 8.209 11.984 −6.991 1.00 33.17 ATOM 1875 CA ASN A 2428.585 13.019 −6.024 1.00 36.07 ATOM 1876 CB ASN A 242 10.059 12.880−5.616 1.00 37.13 ATOM 1877 CG ASN A 242 10.324 11.631 −4.771 1.00 40.96ATOM 1878 OD1 ASN A 242 9.509 11.235 −3.921 1.00 45.33 ATOM 1879 ND2 ASNA 242 11.477 11.007 −4.998 1.00 44.43 ATOM 1880 C ASN A 242 8.321 14.427−6.528 1.00 37.00 ATOM 1881 O ASN A 242 9.091 15.346 −6.245 1.00 37.94ATOM 1882 N GLU A 243 7.210 14.602 −7.233 1.00 37.54 ATOM 1883 CA GLU A243 6.895 15.869 −7.907 1.00 38.05 ATOM 1884 CB GLU A 243 5.775 15.638−8.925 1.00 38.77 ATOM 1885 CG GLU A 243 5.650 16.732 −9.977 1.00 42.65ATOM 1886 CD GLU A 243 6.959 16.985 −10.709 1.00 47.49 ATOM 1887 OE1 GLUA 243 7.424 16.084 −11.453 1.00 49.14 ATOM 1888 OE2 GLU A 243 7.52018.090 −10.532 1.00 50.15 ATOM 1889 C GLU A 243 6.559 17.088 −7.015 1.0037.15 ATOM 1890 O GLU A 243 6.645 18.240 −7.469 1.00 38.39 ATOM 1891 NGLY A 244 6.174 16.873 −5.766 1.00 35.69 ATOM 1892 CA GLY A 244 5.85818.019 −4.911 1.00 33.51 ATOM 1893 C GLY A 244 4.609 18.775 −5.369 1.0031.80 ATOM 1894 O GLY A 244 4.634 19.999 −5.535 1.00 33.32 ATOM 1895 NARG A 245 3.529 18.036 −5.612 1.00 27.92 ATOM 1896 CA ARG A 245 2.20018.618 −5.781 1.00 24.21 ATOM 1897 CB ARG A 245 1.638 18.224 −7.130 1.0024.30 ATOM 1898 CG ARG A 245 2.410 18.842 −8.275 1.00 24.62 ATOM 1899 CDARG A 245 1.625 18.681 −9.532 1.00 22.11 ATOM 1900 NE ARG A 245 2.46218.829 −10.713 1.00 21.13 ATOM 1901 CZ ARG A 245 2.114 18.302 −11.8781.00 21.50 ATOM 1902 NH1 ARG A 245 0.982 17.621 −11.945 1.00 18.57 ATOM1903 NH2 ARG A 245 2.883 18.443 −12.951 1.00 20.83 ATOM 1904 C ARG A 2451.295 18.040 −4.718 1.00 21.84 ATOM 1905 O ARG A 245 1.624 17.021 −4.1281.00 20.65 ATOM 1906 N THR A 246 0.140 18.652 −4.483 1.00 19.15 ATOM1907 CA THR A 246 −0.824 18.058 −3.540 1.00 17.46 ATOM 1908 CB THR A 246−1.989 18.997 −3.238 1.00 17.87 ATOM 1909 OG1 THR A 246 −2.752 19.155−4.440 1.00 15.85 ATOM 1910 CG2 THR A 246 −1.495 20.370 −2.730 1.0017.50 ATOM 1911 C THR A 246 −1.426 16.769 −4.103 1.00 17.25 ATOM 1912 OTHR A 246 −1.884 15.914 −3.351 1.00 17.57 ATOM 1913 N GLY A 247 −1.48216.646 −5.430 1.00 15.60 ATOM 1914 CA GLY A 247 −2.148 15.492 −6.0541.00 15.02 ATOM 1915 C GLY A 247 −3.609 15.761 −6.396 1.00 14.69 ATOM1916 O GLY A 247 −4.260 14.939 −7.059 1.00 14.45 ATOM 1917 N LYS A 248−4.137 16.890 −5.928 1.00 13.43 ATOM 1918 CA LYS A 248 −5.508 17.286−6.259 1.00 13.00 ATOM 1919 CB LYS A 248 −5.969 18.453 −5.396 1.00 12.32ATOM 1920 CG LYS A 248 −5.965 18.179 −3.881 1.00 13.12 ATOM 1921 CD LYSA 248 −6.133 19.493 −3.102 1.00 14.08 ATOM 1922 CE LYS A 248 −5.98519.253 −1.584 1.00 17.84 ATOM 1923 NZ LYS A 248 −6.335 20.492 −0.8351.00 16.74 ATOM 1924 C LYS A 248 −5.490 17.713 −7.736 1.00 12.73 ATOM1925 O LYS A 248 −4.866 18.707 −8.104 1.00 12.75 ATOM 1926 N ASP A 249−6.185 16.964 −8.580 1.00 11.92 ATOM 1927 CA ASP A 249 −5.958 17.098−10.024 1.00 11.16 ATOM 1928 CB ASP A 249 −4.761 16.199 −10.385 1.0010.83 ATOM 1929 CG ASP A 249 −4.268 16.349 −11.831 1.00 12.54 ATOM 1930OD1 ASP A 249 −5.078 16.422 −12.785 1.00 11.42 ATOM 1931 OD2 ASP A 249−3.025 16.342 −12.001 1.00 13.30 ATOM 1932 C ASP A 249 −7.232 16.577−10.662 1.00 11.38 ATOM 1933 O ASP A 249 −7.774 15.542 −10.236 1.0010.86 ATOM 1934 N VAL A 250 −7.700 17.265 −11.703 1.00 11.28 ATOM 1935CA VAL A 250 −8.885 16.793 −12.438 1.00 11.59 ATOM 1936 CB VAL A 250−9.366 17.859 −13.493 1.00 12.49 ATOM 1937 CG1 VAL A 250 −8.480 17.815−14.728 1.00 13.03 ATOM 1938 CG2 VAL A 250 −10.859 17.654 −13.852 1.0013.75 ATOM 1939 C VAL A 250 −8.711 15.386 −13.064 1.00 11.77 ATOM 1940 OVAL A 250 −9.698 14.750 −13.467 1.00 11.71 ATOM 1941 N ASN A 251 −7.46114.925 −13.168 1.00 10.73 ATOM 1942 CA ASN A 251 −7.131 13.491 −13.3781.00 11.20 ATOM 1943 CB ASN A 251 −5.699 13.265 −12.813 1.00 10.94 ATOM1944 CG ASN A 251 −5.221 11.810 −12.892 1.00 11.58 ATOM 1945 OD1 ASN A251 −5.986 10.864 −12.672 1.00 12.47 ATOM 1946 ND2 ASN A 251 −3.89811.639 −13.164 1.00 14.40 ATOM 1947 C ASN A 251 −8.151 12.560 −12.7061.00 10.99 ATOM 1948 O ASN A 251 −8.755 11.706 −13.355 1.00 11.49 ATOM1949 N SER A 252 −8.407 12.774 −11.417 1.00 11.45 ATOM 1950 CA SER A 252−9.293 11.876 −10.634 1.00 11.79 ATOM 1951 CB SER A 252 −9.062 12.155−9.149 1.00 13.31 ATOM 1952 OG SER A 252 −9.338 13.524 −8.882 1.00 13.41ATOM 1953 C SER A 252 −10.784 12.002 −10.996 1.00 11.39 ATOM 1954 O SERA 252 −11.532 11.023 −10.964 1.00 12.69 ATOM 1955 N VAL A 253 −11.19913.203 −11.383 1.00 10.56 ATOM 1956 CA VAL A 253 −12.582 13.459 −11.8211.00 10.70 ATOM 1957 CB VAL A 253 −12.884 15.004 −11.856 1.00 11.02 ATOM1958 CG1 VAL A 253 −14.335 15.262 −12.345 1.00 11.24 ATOM 1959 CG2 VAL A253 −12.711 15.585 −10.449 1.00 10.91 ATOM 1960 C VAL A 253 −12.81012.827 −13.187 1.00 11.38 ATOM 1961 O VAL A 253 −13.824 12.143 −13.4071.00 11.69 ATOM 1962 N LEU A 254 −11.866 13.059 −14.108 1.00 11.32 ATOM1963 CA LEU A 254 −11.891 12.393 −15.417 1.00 12.12 ATOM 1964 CB LEU A254 −10.635 12.759 −16.238 1.00 11.95 ATOM 1965 CG LEU A 254 −10.63414.202 −16.763 1.00 12.23 ATOM 1966 CD1 LEU A 254 −9.266 14.564 −17.3301.00 12.77 ATOM 1967 CD2 LEU A 254 −11.714 14.371 −17.845 1.00 15.26ATOM 1968 C LEU A 254 −11.963 10.872 −15.271 1.00 12.22 ATOM 1969 O LEUA 254 −12.675 10.201 −16.024 1.00 12.01 ATOM 1970 N THR A 255 −11.20810.338 −14.315 1.00 11.58 ATOM 1971 CA THR A 255 −11.219 8.913 −14.0421.00 12.53 ATOM 1972 CB THR A 255 −10.267 8.552 −12.890 1.00 12.83 ATOM1973 OG1 THR A 255 −8.935 8.933 −13.240 1.00 13.00 ATOM 1974 CG2 THR A255 −10.300 7.035 −12.634 1.00 15.06 ATOM 1975 C THR A 255 −12.632 8.448−13.705 1.00 13.12 ATOM 1976 O THR A 255 −13.131 7.467 −14.285 1.0013.49 ATOM 1977 N SER A 256 −13.289 9.158 −12.790 1.00 13.46 ATOM 1978CA SER A 256 −14.641 8.781 −12.343 1.00 12.85 ATOM 1979 CB SER A 256−15.152 9.760 −11.282 1.00 13.40 ATOM 1980 OG SER A 256 −16.332 9.252−10.674 1.00 16.69 ATOM 1981 C SER A 256 −15.610 8.705 −13.518 1.0013.13 ATOM 1982 O SER A 256 −16.360 7.711 −13.654 1.00 13.10 ATOM 1983 NILE A 257 −15.594 9.728 −14.377 1.00 12.32 ATOM 1984 CA ILE A 257−16.523 9.784 −15.513 1.00 12.32 ATOM 1985 CB ILE A 257 −16.747 11.215−16.072 1.00 11.55 ATOM 1986 CG1 ILE A 257 −15.482 11.773 −16.764 1.0011.38 ATOM 1987 CD1 ILE A 257 −15.699 13.143 −17.441 1.00 13.23 ATOM1988 CG2 ILE A 257 −17.257 12.166 −14.942 1.00 13.70 ATOM 1989 C ILE A257 −16.220 8.795 −16.653 1.00 12.79 ATOM 1990 O ILE A 257 −17.150 8.319−17.338 1.00 13.25 ATOM 1991 N HIS A 258 −14.941 8.487 −16.855 1.0012.71 ATOM 1992 CA HIS A 258 −14.565 7.566 −17.931 1.00 13.41 ATOM 1993CB HIS A 258 −13.194 7.947 −18.498 1.00 12.06 ATOM 1994 CG HIS A 258−13.268 9.175 −19.341 1.00 13.92 ATOM 1995 ND1 HIS A 258 −13.942 9.196−20.547 1.00 16.01 ATOM 1996 CE1 HIS A 258 −13.891 10.421 −21.047 1.0018.57 ATOM 1997 NE2 HIS A 258 −13.256 11.199 −20.189 1.00 14.08 ATOM1998 CD2 HIS A 258 −12.861 10.449 −19.108 1.00 13.51 ATOM 1999 C HIS A258 −14.649 6.091 −17.565 1.00 14.12 ATOM 2000 O HIS A 258 −14.645 5.239−18.454 1.00 14.90 ATOM 2001 N THR A 259 −14.752 5.801 −16.274 1.0013.93 ATOM 2002 CA THR A 259 −15.034 4.420 −15.807 1.00 14.91 ATOM 2003CB THR A 259 −13.933 3.856 −14.899 1.00 14.46 ATOM 2004 OG1 THR A 259−13.788 4.647 −13.705 1.00 15.66 ATOM 2005 CG2 THR A 259 −12.589 3.802−15.677 1.00 15.81 ATOM 2006 C THR A 259 −16.433 4.248 −15.173 1.0014.21 ATOM 2007 O THR A 259 −16.709 3.235 −14.546 1.00 14.95 ATOM 2008 NPHE A 260 −17.290 5.238 −15.367 1.00 14.76 ATOM 2009 CA PHE A 260−18.691 5.194 −14.926 1.00 15.13 ATOM 2010 CB PHE A 260 −19.377 6.492−15.379 1.00 15.81 ATOM 2011 CG PHE A 260 −20.886 6.508 −15.228 1.0015.47 ATOM 2012 CD1 PHE A 260 −21.505 6.188 −14.015 1.00 17.59 ATOM 2013CE1 PHE A 260 −22.903 6.259 −13.898 1.00 19.11 ATOM 2014 CZ PHE A 260−23.682 6.653 −14.991 1.00 17.18 ATOM 2015 CE2 PHE A 260 −23.082 6.994−16.178 1.00 18.04 ATOM 2016 CD2 PHE A 260 −21.679 6.917 −16.296 1.0017.22 ATOM 2017 C PHE A 260 −19.436 3.977 −15.475 1.00 15.55 ATOM 2018 OPHE A 260 −19.426 3.725 −16.684 1.00 15.81 ATOM 2019 N ASP A 261 −20.0933.235 −14.586 1.00 15.51 ATOM 2020 CA ASP A 261 −21.008 2.176 −15.0061.00 16.05 ATOM 2021 CB ASP A 261 −20.303 0.813 −15.015 1.00 16.46 ATOM2022 CG ASP A 261 −21.205 −0.321 −15.490 1.00 17.60 ATOM 2023 OD1 ASP A261 −22.440 −0.122 −15.579 1.00 18.97 ATOM 2024 OD2 ASP A 261 −20.656−1.404 −15.810 1.00 18.29 ATOM 2025 C ASP A 261 −22.117 2.185 −13.9721.00 16.30 ATOM 2026 O ASP A 261 −21.882 1.809 −12.840 1.00 15.53 ATOM2027 N PRO A 262 −23.320 2.610 −14.374 1.00 18.21 ATOM 2028 CA PRO A 262−24.438 2.716 −13.412 1.00 20.39 ATOM 2029 CB PRO A 262 −25.589 3.308−14.247 1.00 20.43 ATOM 2030 CG PRO A 262 −25.235 3.044 −15.669 1.0020.06 ATOM 2031 CD PRO A 262 −23.709 2.994 −15.734 1.00 17.40 ATOM 2032C PRO A 262 −24.815 1.382 −12.753 1.00 22.31 ATOM 2033 O PRO A 262−25.356 1.374 −11.622 1.00 22.24 ATOM 2034 N ASN A 263 −24.508 0.267−13.421 1.00 22.99 ATOM 2035 CA ASN A 263 −24.750 −1.048 −12.838 1.0025.42 ATOM 2036 CB ASN A 263 −24.574 −2.149 −13.890 1.00 26.62 ATOM 2037CG ASN A 263 −25.680 −2.128 −14.948 1.00 30.74 ATOM 2038 OD1 ASN A 263−26.688 −1.419 −14.814 1.00 35.92 ATOM 2039 ND2 ASN A 263 −25.490 −2.906−16.007 1.00 35.87 ATOM 2040 C ASN A 263 −23.894 −1.316 −11.598 1.0025.45 ATOM 2041 O ASN A 263 −24.210 −2.190 −10.795 1.00 26.56 ATOM 2042N LEU A 264 −22.835 −0.529 −11.413 1.00 24.54 ATOM 2043 CA LEU A 264−22.022 −0.616 −10.213 1.00 24.27 ATOM 2044 CB LEU A 264 −20.549 −0.287−10.520 1.00 24.43 ATOM 2045 CG LEU A 264 −19.752 −1.346 −11.288 1.0025.38 ATOM 2046 CD1 LEU A 264 −18.375 −0.809 −11.659 1.00 26.05 ATOM2047 CD2 LEU A 264 −19.619 −2.672 −10.523 1.00 26.24 ATOM 2048 C LEU A264 −22.542 0.273 −9.066 1.00 23.47 ATOM 2049 O LEU A 264 −21.956 0.292−7.988 1.00 23.97 ATOM 2050 N GLY A 265 −23.631 1.000 −9.292 1.00 23.32ATOM 2051 CA GLY A 265 −24.218 1.840 −8.237 1.00 22.74 ATOM 2052 C GLY A265 −23.204 2.843 −7.729 1.00 21.84 ATOM 2053 O GLY A 265 −22.416 3.373−8.510 1.00 22.83 ATOM 2054 N CYS A 266 −23.175 3.086 −6.424 1.00 21.37ATOM 2055 CA CYS A 266 −22.233 4.073 −5.883 1.00 21.00 ATOM 2056 CB CYSA 266 −22.947 5.049 −4.936 1.00 20.86 ATOM 2057 SG CYS A 266 −24.3475.912 −5.711 1.00 20.96 ATOM 2058 C CYS A 266 −20.992 3.427 −5.275 1.0020.98 ATOM 2059 O CYS A 266 −20.513 3.814 −4.203 1.00 21.01 ATOM 2060 NASP A 267 −20.462 2.443 −6.002 1.00 20.39 ATOM 2061 CA ASP A 267 −19.3031.686 −5.577 1.00 20.67 ATOM 2062 CB ASP A 267 −18.961 0.621 −6.618 1.0020.84 ATOM 2063 CG ASP A 267 −17.666 −0.101 −6.288 1.00 24.28 ATOM 2064OD1 ASP A 267 −16.852 −0.322 −7.200 1.00 25.84 ATOM 2065 OD2 ASP A 267−17.455 −0.407 −5.098 1.00 27.46 ATOM 2066 C ASP A 267 −18.072 2.567−5.391 1.00 19.78 ATOM 2067 O ASP A 267 −17.593 3.161 −6.353 1.00 18.61ATOM 2068 N ALA A 268 −17.544 2.621 −4.174 1.00 18.35 ATOM 2069 CA ALA A268 −16.315 3.395 −3.944 1.00 19.35 ATOM 2070 CB ALA A 268 −16.207 3.868−2.472 1.00 19.46 ATOM 2071 C ALA A 268 −15.017 2.701 −4.415 1.00 19.46ATOM 2072 O ALA A 268 −14.009 3.371 −4.665 1.00 19.42 ATOM 2073 N GLY A269 −15.029 1.370 −4.534 1.00 19.01 ATOM 2074 CA GLY A 269 −13.826 0.644−4.936 1.00 19.17 ATOM 2075 C GLY A 269 −13.370 1.032 −6.343 1.00 18.75ATOM 2076 O GLY A 269 −12.175 1.134 −6.624 1.00 19.54 ATOM 2077 N THR A270 −14.330 1.257 −7.230 1.00 17.92 ATOM 2078 CA THR A 270 −14.016 1.662−8.594 1.00 18.35 ATOM 2079 CB THR A 270 −14.852 0.882 −9.616 1.00 18.43ATOM 2080 OG1 THR A 270 −16.246 1.085 −9.350 1.00 18.51 ATOM 2081 CG2THR A 270 −14.529 −0.626 −9.555 1.00 19.75 ATOM 2082 C THR A 270 −14.2613.172 −8.771 1.00 18.25 ATOM 2083 O THR A 270 −14.326 3.674 −9.904 1.0017.88 ATOM 2084 N PHE A 271 −14.434 3.880 −7.650 1.00 17.17 ATOM 2085 CAPHE A 271 −14.531 5.359 −7.656 1.00 17.38 ATOM 2086 CB PHE A 271 −13.1835.965 −8.121 1.00 17.67 ATOM 2087 CG PHE A 271 −12.946 7.376 −7.673 1.0021.97 ATOM 2088 CD1 PHE A 271 −12.656 7.653 −6.337 1.00 24.90 ATOM 2089CE1 PHE A 271 −12.447 8.981 −5.923 1.00 24.46 ATOM 2090 CZ PHE A 271−12.474 10.043 −6.863 1.00 23.20 ATOM 2091 CE2 PHE A 271 −12.733 9.783−8.196 1.00 21.92 ATOM 2092 CD2 PHE A 271 −12.956 8.436 −8.599 1.0024.30 ATOM 2093 C PHE A 271 −15.677 5.856 −8.551 1.00 16.63 ATOM 2094 OPHE A 271 −15.479 6.764 −9.358 1.00 15.93 ATOM 2095 N GLN A 272 −16.8615.249 −8.439 1.00 15.21 ATOM 2096 CA GLN A 272 −18.011 5.673 −9.251 1.0014.97 ATOM 2097 CB GLN A 272 −19.227 4.755 −9.013 1.00 14.93 ATOM 2098CG GLN A 272 −19.021 3.355 −9.615 1.00 16.30 ATOM 2099 CD GLN A 272−18.755 3.413 −11.102 1.00 15.81 ATOM 2100 OE1 GLN A 272 −19.575 3.909−11.883 1.00 16.97 ATOM 2101 NE2 GLN A 272 −17.617 2.861 −11.512 1.0018.50 ATOM 2102 C GLN A 272 −18.402 7.118 −8.929 1.00 14.73 ATOM 2103 OGLN A 272 −18.194 7.555 −7.800 1.00 15.60 ATOM 2104 N PRO A 273 −18.9557.859 −9.914 1.00 14.45 ATOM 2105 CA PRO A 273 −19.342 9.255 −9.682 1.0014.57 ATOM 2106 CB PRO A 273 −20.157 9.597 −10.927 1.00 14.64 ATOM 2107CG PRO A 273 −19.443 8.767 −12.031 1.00 14.70 ATOM 2108 CD PRO A 273−19.156 7.458 −11.326 1.00 14.10 ATOM 2109 C PRO A 273 −20.162 9.542−8.407 1.00 15.22 ATOM 2110 O PRO A 273 −19.910 10.562 −7.752 1.00 15.03ATOM 2111 N CYS A 274 −21.130 8.682 −8.075 1.00 15.76 ATOM 2112 CA CYS A274 −21.926 8.913 −6.853 1.00 16.22 ATOM 2113 CB CYS A 274 −23.389 8.489−7.039 1.00 16.57 ATOM 2114 SG CYS A 274 −23.611 6.769 −7.423 1.00 17.39ATOM 2115 C CYS A 274 −21.331 8.281 −5.605 1.00 16.64 ATOM 2116 O CYS A274 −21.958 8.329 −4.529 1.00 16.55 ATOM 2117 N SER A 275 −20.137 7.681−5.715 1.00 15.61 ATOM 2118 CA SER A 275 −19.476 7.117 −4.528 1.00 15.81ATOM 2119 CB SER A 275 −18.244 6.253 −4.877 1.00 15.06 ATOM 2120 OG SERA 275 −17.144 7.041 −5.315 1.00 14.92 ATOM 2121 C SER A 275 −19.0978.232 −3.545 1.00 16.06 ATOM 2122 O SER A 275 −18.818 9.366 −3.949 1.0014.39 ATOM 2123 N ASP A 276 −19.103 7.919 −2.248 1.00 16.42 ATOM 2124 CAASP A 276 −18.731 8.935 −1.271 1.00 16.52 ATOM 2125 CB ASP A 276 −19.0208.511 0.189 1.00 16.27 ATOM 2126 CG ASP A 276 −18.244 7.281 0.656 1.0019.14 ATOM 2127 OD1 ASP A 276 −18.371 7.001 1.873 1.00 19.62 ATOM 2128OD2 ASP A 276 −17.544 6.593 −0.120 1.00 17.10 ATOM 2129 C ASP A 276−17.312 9.469 −1.492 1.00 16.17 ATOM 2130 O ASP A 276 −17.084 10.683−1.415 1.00 15.20 ATOM 2131 N LYS A 277 −16.381 8.577 −1.823 1.00 15.43ATOM 2132 CA LYS A 277 −14.994 8.982 −2.115 1.00 15.34 ATOM 2133 CB LYSA 277 −14.089 7.763 −2.326 1.00 15.23 ATOM 2134 CG LYS A 277 −13.9246.905 −1.059 1.00 17.01 ATOM 2135 CD LYS A 277 −12.752 5.929 −1.204 1.0021.20 ATOM 2136 CE LYS A 277 −12.662 5.017 0.015 1.00 22.94 ATOM 2137 NZLYS A 277 −11.533 4.067 −0.165 1.00 29.19 ATOM 2138 C LYS A 277 −14.9009.915 −3.324 1.00 14.30 ATOM 2139 O LYS A 277 −14.152 10.887 −3.288 1.0014.70 ATOM 2140 N ALA A 278 −15.644 9.620 −4.393 1.00 14.45 ATOM 2141 CAALA A 278 −15.588 10.464 −5.605 1.00 13.61 ATOM 2142 CB ALA A 278−16.250 9.775 −6.783 1.00 13.30 ATOM 2143 C ALA A 278 −16.210 11.827−5.357 1.00 13.50 ATOM 2144 O ALA A 278 −15.730 12.840 −5.864 1.00 13.02ATOM 2145 N LEU A 279 −17.283 11.855 −4.565 1.00 13.22 ATOM 2146 CA LEUA 279 −17.936 13.132 −4.239 1.00 12.92 ATOM 2147 CB LEU A 279 −19.32312.893 −3.625 1.00 13.21 ATOM 2148 CG LEU A 279 −20.384 12.358 −4.6011.00 13.94 ATOM 2149 CD1 LEU A 279 −21.707 11.969 −3.887 1.00 17.68 ATOM2150 CD2 LEU A 279 −20.653 13.319 −5.781 1.00 17.52 ATOM 2151 C LEU A279 −17.065 13.995 −3.348 1.00 12.84 ATOM 2152 O LEU A 279 −16.94115.203 −3.577 1.00 13.54 ATOM 2153 N SER A 280 −16.463 13.390 −2.3151.00 12.45 ATOM 2154 CA SER A 280 −15.502 14.106 −1.459 1.00 13.53 ATOM2155 CB SER A 280 −14.951 13.168 −0.364 1.00 13.65 ATOM 2156 OG SER A280 −14.008 13.863 0.468 1.00 15.07 ATOM 2157 C SER A 280 −14.332 14.672−2.285 1.00 14.09 ATOM 2158 O SER A 280 −13.925 15.856 −2.130 1.00 13.43ATOM 2159 N ASN A 281 −13.795 13.830 −3.166 1.00 13.27 ATOM 2160 CA ASNA 281 −12.690 14.257 −4.027 1.00 13.19 ATOM 2161 CB ASN A 281 −12.23913.078 −4.888 1.00 12.05 ATOM 2162 CG ASN A 281 −11.116 13.455 −5.8491.00 13.13 ATOM 2163 OD1 ASN A 281 −9.989 13.637 −5.446 1.00 13.26 ATOM2164 ND2 ASN A 281 −11.442 13.573 −7.124 1.00 11.63 ATOM 2165 C ASN A281 −13.096 15.432 −4.933 1.00 12.33 ATOM 2166 O ASN A 281 −12.33016.380 −5.109 1.00 13.49 ATOM 2167 N LEU A 282 −14.287 15.355 −5.5061.00 12.05 ATOM 2168 CA LEU A 282 −14.760 16.422 −6.376 1.00 13.21 ATOM2169 CB LEU A 282 −16.147 16.109 −6.949 1.00 12.17 ATOM 2170 CG LEU A282 −16.791 17.216 −7.820 1.00 14.57 ATOM 2171 CD1 LEU A 282 −16.01117.378 −9.126 1.00 16.58 ATOM 2172 CD2 LEU A 282 −18.241 16.863 −8.1701.00 15.68 ATOM 2173 C LEU A 282 −14.739 17.754 −5.638 1.00 12.69 ATOM2174 O LEU A 282 −14.201 18.735 −6.153 1.00 13.45 ATOM 2175 N LYS A 283−15.283 17.791 −4.415 1.00 12.75 ATOM 2176 CA LYS A 283 −15.306 19.026−3.656 1.00 12.89 ATOM 2177 CB LYS A 283 −16.079 18.860 −2.334 1.0012.90 ATOM 2178 CG LYS A 283 −15.912 20.089 −1.432 1.00 13.94 ATOM 2179CD LYS A 283 −16.909 20.076 −0.252 1.00 14.67 ATOM 2180 CE LYS A 283−16.530 21.136 0.797 1.00 13.67 ATOM 2181 NZ LYS A 283 −16.315 22.4890.212 1.00 19.03 ATOM 2182 C LYS A 283 −13.889 19.537 −3.385 1.00 12.43ATOM 2183 O LYS A 283 −13.612 20.710 −3.556 1.00 12.14 ATOM 2184 N VAL A284 −12.988 18.652 −2.966 1.00 12.02 ATOM 2185 CA VAL A 284 −11.62419.055 −2.633 1.00 12.77 ATOM 2186 CB VAL A 284 −10.845 17.875 −2.0141.00 13.17 ATOM 2187 CG1 VAL A 284 −9.320 18.169 −1.936 1.00 13.21 ATOM2188 CG2 VAL A 284 −11.391 17.557 −0.630 1.00 15.81 ATOM 2189 C VAL A284 −10.927 19.599 −3.881 1.00 12.74 ATOM 2190 O VAL A 284 −10.22820.636 −3.827 1.00 12.21 ATOM 2191 N VAL A 285 −11.153 18.927 −5.0121.00 11.54 ATOM 2192 CA VAL A 285 −10.560 19.389 −6.287 1.00 12.35 ATOM2193 CB VAL A 285 −10.694 18.330 −7.425 1.00 12.36 ATOM 2194 CG1 VAL A285 −10.316 18.944 −8.813 1.00 12.25 ATOM 2195 CG2 VAL A 285 −9.79517.104 −7.140 1.00 13.25 ATOM 2196 C VAL A 285 −11.130 20.770 −6.7121.00 12.08 ATOM 2197 O VAL A 285 −10.367 21.696 −6.989 1.00 12.60 ATOM2198 N VAL A 286 −12.452 20.913 −6.728 1.00 11.87 ATOM 2199 CA VAL A 286−13.089 22.196 −7.074 1.00 12.87 ATOM 2200 CB VAL A 286 −14.631 22.080−7.038 1.00 13.01 ATOM 2201 CG1 VAL A 286 −15.300 23.468 −7.140 1.0014.31 ATOM 2202 CG2 VAL A 286 −15.103 21.157 −8.200 1.00 14.42 ATOM 2203C VAL A 286 −12.586 23.324 −6.164 1.00 12.84 ATOM 2204 O VAL A 286−12.206 24.402 −6.635 1.00 13.75 ATOM 2205 N ASP A 287 −12.552 23.064−4.853 1.00 12.85 ATOM 2206 CA ASP A 287 −12.116 24.059 −3.870 1.0013.90 ATOM 2207 CB ASP A 287 −12.199 23.506 −2.440 1.00 13.12 ATOM 2208CG ASP A 287 −13.637 23.441 −1.924 1.00 16.00 ATOM 2209 OD1 ASP A 287−14.541 24.002 −2.583 1.00 16.20 ATOM 2210 OD2 ASP A 287 −13.857 22.835−0.858 1.00 16.76 ATOM 2211 C ASP A 287 −10.727 24.564 −4.136 1.00 14.28ATOM 2212 O ASP A 287 −10.425 25.722 −3.841 1.00 15.53 ATOM 2213 N SER A288 −9.862 23.709 −4.677 1.00 14.50 ATOM 2214 CA SER A 288 −8.478 24.093−4.949 1.00 14.58 ATOM 2215 CB SER A 288 −7.625 22.843 −5.229 1.00 14.15ATOM 2216 OG SER A 288 −7.758 22.417 −6.565 1.00 13.73 ATOM 2217 C SER A288 −8.326 25.186 −6.038 1.00 14.61 ATOM 2218 O SER A 288 −7.274 25.847−6.143 1.00 14.59 ATOM 2219 N PHE A 289 −9.392 25.416 −6.809 1.00 13.99ATOM 2220 CA PHE A 289 −9.419 26.447 −7.831 1.00 14.09 ATOM 2221 CB PHEA 289 −9.994 25.882 −9.135 1.00 13.52 ATOM 2222 CG PHE A 289 −9.16924.807 −9.704 1.00 11.38 ATOM 2223 CD1 PHE A 289 −7.976 25.114 −10.3671.00 12.17 ATOM 2224 CE1 PHE A 289 −7.184 24.095 −10.905 1.00 13.93 ATOM2225 CZ PHE A 289 −7.572 22.783 −10.771 1.00 13.96 ATOM 2226 CE2 PHE A289 −8.756 22.452 −10.097 1.00 11.77 ATOM 2227 CD2 PHE A 289 −9.55523.472 −9.571 1.00 11.08 ATOM 2228 C PHE A 289 −10.219 27.698 −7.4911.00 14.73 ATOM 2229 O PHE A 289 −10.092 28.713 −8.189 1.00 14.71 ATOM2230 N ARG A 290 −11.054 27.621 −6.464 1.00 15.09 ATOM 2231 CA ARG A 290−11.953 28.740 −6.140 1.00 16.64 ATOM 2232 CB ARG A 290 −12.842 28.401−4.936 1.00 15.99 ATOM 2233 CG ARG A 290 −13.913 27.375 −5.230 1.0015.65 ATOM 2234 CD ARG A 290 −14.821 27.163 −4.012 1.00 16.79 ATOM 2235NE ARG A 290 −15.843 26.172 −4.330 1.00 15.04 ATOM 2236 CZ ARG A 290−16.986 26.470 −4.933 1.00 17.22 ATOM 2237 NH1 ARG A 290 −17.248 27.734−5.243 1.00 15.41 ATOM 2238 NH2 ARG A 290 −17.855 25.511 −5.239 1.0015.94 ATOM 2239 C ARG A 290 −11.240 30.046 −5.864 1.00 18.04 ATOM 2240 OARG A 290 −11.690 31.125 −6.279 1.00 19.61 ATOM 2241 N SER A 291 −10.15029.984 −5.128 1.00 19.44 ATOM 2242 CA SER A 291 −9.571 31.246 −4.6671.00 21.57 ATOM 2243 CB SER A 291 −9.146 31.101 −3.212 1.00 22.18 ATOM2244 OG SER A 291 −7.998 30.284 −3.144 1.00 28.35 ATOM 2245 C SER A 291−8.423 31.762 −5.534 1.00 20.65 ATOM 2246 O SER A 291 −7.865 32.851−5.272 1.00 22.43 ATOM 2247 N ILE A 292 −8.066 31.019 −6.576 1.00 19.16ATOM 2248 CA ILE A 292 −6.855 31.367 −7.330 1.00 17.65 ATOM 2249 CB ILEA 292 −5.805 30.185 −7.408 1.00 18.07 ATOM 2250 CG1 ILE A 292 −6.37928.972 −8.194 1.00 17.67 ATOM 2251 CD1 ILE A 292 −5.315 27.924 −8.6491.00 17.27 ATOM 2252 CG2 ILE A 292 −5.341 29.795 −5.994 1.00 18.29 ATOM2253 C ILE A 292 −7.065 31.973 −8.708 1.00 17.20 ATOM 2254 O ILE A 292−6.136 32.563 −9.251 1.00 16.35 ATOM 2255 N TYR A 293 −8.252 31.797−9.290 1.00 15.85 ATOM 2256 CA TYR A 293 −8.509 32.304 −10.648 1.0015.83 ATOM 2257 CB TYR A 293 −9.301 31.270 −11.474 1.00 15.43 ATOM 2258CG TYR A 293 −8.571 30.014 −11.886 1.00 15.10 ATOM 2259 CD1 TYR A 293−7.183 29.960 −11.965 1.00 14.38 ATOM 2260 CE1 TYR A 293 −6.540 28.795−12.395 1.00 14.38 ATOM 2261 CZ TYR A 293 −7.306 27.685 −12.743 1.0014.90 ATOM 2262 OH TYR A 293 −6.700 26.522 −13.158 1.00 15.55 ATOM 2263CE2 TYR A 293 −8.670 27.722 −12.671 1.00 15.47 ATOM 2264 CD2 TYR A 293−9.298 28.875 −12.255 1.00 13.91 ATOM 2265 C TYR A 293 −9.351 33.581−10.591 1.00 15.69 ATOM 2266 O TYR A 293 −10.404 33.594 −9.942 1.0015.47 ATOM 2267 N GLY A 294 −8.892 34.629 −11.276 1.00 14.83 ATOM 2268CA GLY A 294 −9.641 35.899 −11.353 1.00 15.57 ATOM 2269 C GLY A 294−11.078 35.702 −11.858 1.00 15.93 ATOM 2270 O GLY A 294 −12.010 36.359−11.376 1.00 15.66 ATOM 2271 N VAL A 295 −11.288 34.773 −12.799 1.0015.60 ATOM 2272 CA VAL A 295 −12.651 34.520 −13.270 1.00 16.24 ATOM 2273CB VAL A 295 −12.753 33.561 −14.501 1.00 16.31 ATOM 2274 CG1 VAL A 295−12.170 34.195 −15.740 1.00 16.26 ATOM 2275 CG2 VAL A 295 −12.128 32.184−14.199 1.00 16.19 ATOM 2276 C VAL A 295 −13.596 34.013 −12.172 1.0016.97 ATOM 2277 O VAL A 295 −14.813 34.108 −12.320 1.00 18.03 ATOM 2278N ASN A 296 −13.047 33.463 −11.092 1.00 16.93 ATOM 2279 CA ASN A 296−13.878 32.920 −10.020 1.00 17.69 ATOM 2280 CB ASN A 296 −13.250 31.633−9.472 1.00 17.57 ATOM 2281 CG ASN A 296 −13.296 30.493 −10.482 1.0016.44 ATOM 2282 OD1 ASN A 296 −14.158 30.481 −11.356 1.00 17.29 ATOM2283 ND2 ASN A 296 −12.401 29.513 −10.336 1.00 15.99 ATOM 2284 C ASN A296 −14.187 33.915 −8.896 1.00 19.30 ATOM 2285 O ASN A 296 −14.94533.601 −7.979 1.00 19.07 ATOM 2286 N LYS A 297 −13.617 35.116 −9.0071.00 20.37 ATOM 2287 CA LYS A 297 −13.811 36.203 −8.038 1.00 22.43 ATOM2288 CB LYS A 297 −13.209 37.502 −8.584 1.00 22.90 ATOM 2289 CG LYS A297 −11.741 37.680 −8.316 1.00 30.03 ATOM 2290 CD LYS A 297 −11.40139.189 −8.309 1.00 35.34 ATOM 2291 CE LYS A 297 −12.247 39.913 −7.2551.00 39.86 ATOM 2292 NZ LYS A 297 −11.995 41.386 −7.178 1.00 42.72 ATOM2293 C LYS A 297 −15.275 36.453 −7.782 1.00 21.89 ATOM 2294 O LYS A 297−16.061 36.585 −8.712 1.00 21.96 ATOM 2295 N GLY A 298 −15.659 36.537−6.517 1.00 22.72 ATOM 2296 CA GLY A 298 −17.050 36.869 −6.219 1.0022.99 ATOM 2297 C GLY A 298 −18.043 35.720 −6.278 1.00 23.54 ATOM 2298 OGLY A 298 −19.180 35.885 −5.855 1.00 25.04 ATOM 2299 N ILE A 299 −17.64734.546 −6.784 1.00 22.16 ATOM 2300 CA ILE A 299 −18.574 33.393 −6.7631.00 21.47 ATOM 2301 CB ILE A 299 −18.251 32.350 −7.884 1.00 21.22 ATOM2302 CG1 ILE A 299 −18.356 32.985 −9.274 1.00 19.64 ATOM 2303 CD1 ILE A299 −17.740 32.095 −10.415 1.00 19.76 ATOM 2304 CG2 ILE A 299 −19.16331.091 −7.762 1.00 20.54 ATOM 2305 C ILE A 299 −18.562 32.740 −5.3751.00 22.34 ATOM 2306 O ILE A 299 −17.486 32.395 −4.861 1.00 22.29 ATOM2307 N PRO A 300 −19.743 32.580 −4.751 1.00 23.04 ATOM 2308 CA PRO A 300−19.791 32.018 −3.392 1.00 23.60 ATOM 2309 CB PRO A 300 −21.217 32.364−2.922 1.00 24.19 ATOM 2310 CG PRO A 300 −22.015 32.437 −4.178 1.0023.80 ATOM 2311 CD PRO A 300 −21.085 32.934 −5.253 1.00 23.06 ATOM 2312C PRO A 300 −19.584 30.500 −3.322 1.00 23.50 ATOM 2313 O PRO A 300−19.664 29.810 −4.347 1.00 22.46 ATOM 2314 N ALA A 301 −19.325 29.985−2.116 1.00 22.68 ATOM 2315 CA ALA A 301 −19.380 28.549 −1.905 1.0022.89 ATOM 2316 CB ALA A 301 −18.988 28.185 −0.465 1.00 23.54 ATOM 2317C ALA A 301 −20.788 28.074 −2.236 1.00 21.91 ATOM 2318 O ALA A 301−21.759 28.834 −2.108 1.00 23.09 ATOM 2319 N GLY A 302 −20.898 26.838−2.698 1.00 20.96 ATOM 2320 CA GLY A 302 −22.173 26.272 −3.115 1.0019.69 ATOM 2321 C GLY A 302 −22.565 26.637 −4.537 1.00 19.78 ATOM 2322 OGLY A 302 −23.661 26.283 −4.991 1.00 19.32 ATOM 2323 N ALA A 303 −21.68627.355 −5.235 1.00 17.97 ATOM 2324 CA ALA A 303 −21.948 27.708 −6.6351.00 17.13 ATOM 2325 CB ALA A 303 −22.168 29.212 −6.812 1.00 16.73 ATOM2326 C ALA A 303 −20.784 27.245 −7.481 1.00 16.19 ATOM 2327 O ALA A 303−19.647 27.171 −7.004 1.00 16.23 ATOM 2328 N ALA A 304 −21.067 26.956−8.746 1.00 15.66 ATOM 2329 CA ALA A 304 −20.069 26.378 −9.640 1.0015.10 ATOM 2330 CB ALA A 304 −20.750 25.795 −10.860 1.00 15.80 ATOM 2331C ALA A 304 −19.002 27.394 −10.044 1.00 14.74 ATOM 2332 O ALA A 304−19.270 28.587 −10.121 1.00 14.27 ATOM 2333 N VAL A 305 −17.783 26.914−10.300 1.00 14.18 ATOM 2334 CA VAL A 305 −16.680 27.783 −10.698 1.0013.54 ATOM 2335 CB VAL A 305 −15.656 27.971 −9.543 1.00 13.42 ATOM 2336CG1 VAL A 305 −16.224 28.881 −8.418 1.00 14.25 ATOM 2337 CG2 VAL A 305−15.218 26.597 −8.966 1.00 14.99 ATOM 2338 C VAL A 305 −15.952 27.141−11.873 1.00 13.34 ATOM 2339 O VAL A 305 −16.121 25.944 −12.126 1.0012.46 ATOM 2340 N ALA A 306 −15.130 27.921 −12.562 1.00 13.38 ATOM 2341CA ALA A 306 −14.233 27.376 −13.573 1.00 14.37 ATOM 2342 CB ALA A 306−13.709 28.504 −14.470 1.00 15.32 ATOM 2343 C ALA A 306 −13.082 26.626−12.938 1.00 14.68 ATOM 2344 O ALA A 306 −12.457 27.116 −11.974 1.0015.21 ATOM 2345 N ILE A 307 −12.781 25.452 −13.484 1.00 13.50 ATOM 2346CA ILE A 307 −11.667 24.668 −12.975 1.00 13.67 ATOM 2347 CB ILE A 307−12.134 23.438 −12.163 1.00 14.45 ATOM 2348 CG1 ILE A 307 −12.756 22.386−13.072 1.00 15.37 ATOM 2349 CD1 ILE A 307 −12.921 21.033 −12.368 1.0019.15 ATOM 2350 CG2 ILE A 307 −13.119 23.848 −11.005 1.00 15.52 ATOM2351 C ILE A 307 −10.646 24.290 −14.059 1.00 12.52 ATOM 2352 O ILE A 307−10.974 24.232 −15.264 1.00 12.22 ATOM 2353 N GLY A 308 −9.405 24.095−13.604 1.00 11.71 ATOM 2354 CA GLY A 308 −8.276 23.737 −14.452 1.0011.64 ATOM 2355 C GLY A 308 −7.853 22.306 −14.199 1.00 11.62 ATOM 2356 OGLY A 308 −8.667 21.444 −13.806 1.00 12.05 ATOM 2357 N ARG A 309 −6.58322.026 −14.454 1.00 11.39 ATOM 2358 CA ARG A 309 −6.091 20.661 −14.3371.00 11.23 ATOM 2359 CB ARG A 309 −4.896 20.467 −15.275 1.00 11.37 ATOM2360 CG ARG A 309 −5.220 20.697 −16.791 1.00 11.29 ATOM 2361 CD ARG A309 −4.066 20.130 −17.625 1.00 12.62 ATOM 2362 NE ARG A 309 −2.84520.919 −17.425 1.00 12.15 ATOM 2363 CZ ARG A 309 −1.701 20.665 −18.0471.00 15.00 ATOM 2364 NH1 ARG A 309 −1.630 19.633 −18.910 1.00 12.05 ATOM2365 NH2 ARG A 309 −0.624 21.395 −17.778 1.00 13.85 ATOM 2366 C ARG A309 −5.654 20.425 −12.888 1.00 11.83 ATOM 2367 O ARG A 309 −6.093 19.481−12.221 1.00 11.38 ATOM 2368 N TYR A 310 −4.806 21.322 −12.399 1.0011.88 ATOM 2369 CA TYR A 310 −4.293 21.215 −11.022 1.00 11.17 ATOM 2370CB TYR A 310 −3.225 20.082 −10.878 1.00 12.49 ATOM 2371 CG TYR A 310−2.065 20.201 −11.844 1.00 13.10 ATOM 2372 CD1 TYR A 310 −2.128 19.622−13.138 1.00 12.57 ATOM 2373 CE1 TYR A 310 −1.069 19.772 −14.039 1.0015.63 ATOM 2374 CZ TYR A 310 0.065 20.475 −13.649 1.00 14.31 ATOM 2375OH TYR A 310 1.119 20.611 −14.529 1.00 14.42 ATOM 2376 CE2 TYR A 3100.159 21.030 −12.379 1.00 12.68 ATOM 2377 CD2 TYR A 310 −0.909 20.906−11.485 1.00 14.04 ATOM 2378 C TYR A 310 −3.779 22.596 −10.644 1.0012.74 ATOM 2379 O TYR A 310 −3.333 23.356 −11.505 1.00 12.44 ATOM 2380 NALA A 311 −3.872 22.945 −9.362 1.00 11.99 ATOM 2381 CA ALA A 311 −3.61824.337 −8.975 1.00 13.33 ATOM 2382 CB ALA A 311 −4.084 24.589 −7.5081.00 12.88 ATOM 2383 C ALA A 311 −2.157 24.768 −9.197 1.00 13.50 ATOM2384 O ALA A 311 −1.906 25.951 −9.468 1.00 14.52 ATOM 2385 N GLU A 312−1.216 23.823 −9.140 1.00 13.52 ATOM 2386 CA GLU A 312 0.219 24.134−9.332 1.00 14.23 ATOM 2387 CB GLU A 312 1.111 23.020 −8.790 1.00 15.44ATOM 2388 CG GLU A 312 0.933 22.802 −7.303 1.00 16.54 ATOM 2389 CD GLU A312 −0.130 21.762 −6.950 1.00 18.91 ATOM 2390 OE1 GLU A 312 −0.94121.338 −7.808 1.00 16.89 ATOM 2391 OE2 GLU A 312 −0.150 21.345 −5.7781.00 18.72 ATOM 2392 C GLU A 312 0.591 24.380 −10.796 1.00 14.80 ATOM2393 O GLU A 312 1.741 24.736 −11.100 1.00 14.98 ATOM 2394 N ASP A 313−0.374 24.197 −11.697 1.00 13.37 ATOM 2395 CA ASP A 313 −0.112 24.258−13.155 1.00 13.71 ATOM 2396 CB ASP A 313 −1.457 24.079 −13.888 1.0012.88 ATOM 2397 CG ASP A 313 −1.320 23.671 −15.343 1.00 14.44 ATOM 2398OD1 ASP A 313 −0.197 23.597 −15.900 1.00 13.19 ATOM 2399 OD2 ASP A 313−2.400 23.406 −15.923 1.00 13.61 ATOM 2400 C ASP A 313 0.512 25.589−13.587 1.00 14.00 ATOM 2401 O ASP A 313 0.007 26.662 −13.219 1.00 14.35ATOM 2402 N VAL A 314 1.577 25.530 −14.399 1.00 13.84 ATOM 2403 CA VAL A314 2.145 26.747 −14.988 1.00 15.19 ATOM 2404 CB VAL A 314 3.602 27.016−14.520 1.00 16.70 ATOM 2405 CG1 VAL A 314 3.638 27.295 −13.009 1.0017.94 ATOM 2406 CG2 VAL A 314 4.551 25.857 −14.915 1.00 16.69 ATOM 2407C VAL A 314 2.123 26.729 −16.528 1.00 15.30 ATOM 2408 O VAL A 314 2.71227.598 −17.165 1.00 15.13 ATOM 2409 N TYR A 315 1.441 25.743 −17.1111.00 14.51 ATOM 2410 CA TYR A 315 1.351 25.634 −18.580 1.00 15.65 ATOM2411 CB TYR A 315 0.768 24.264 −18.957 1.00 15.82 ATOM 2412 CG TYR A 3150.694 23.988 −20.457 1.00 16.29 ATOM 2413 CD1 TYR A 315 1.824 24.124−21.265 1.00 17.01 ATOM 2414 CE1 TYR A 315 1.778 23.859 −22.634 1.0018.92 ATOM 2415 CZ TYR A 315 0.588 23.421 −23.208 1.00 16.14 ATOM 2416OH TYR A 315 0.557 23.164 −24.577 1.00 16.95 ATOM 2417 CE2 TYR A 315−0.552 23.261 −22.423 1.00 15.41 ATOM 2418 CD2 TYR A 315 −0.492 23.539−21.044 1.00 14.48 ATOM 2419 C TYR A 315 0.489 26.777 −19.107 1.00 15.56ATOM 2420 O TYR A 315 −0.688 26.888 −18.748 1.00 16.36 ATOM 2421 N TYR A316 1.072 27.645 −19.944 1.00 16.61 ATOM 2422 CA TYR A 316 0.404 28.890−20.380 1.00 17.53 ATOM 2423 CB TYR A 316 −0.778 28.603 −21.337 1.0018.31 ATOM 2424 CG TYR A 316 −0.329 28.321 −22.742 1.00 19.74 ATOM 2425CD1 TYR A 316 −0.071 27.026 −23.169 1.00 18.95 ATOM 2426 CE1 TYR A 3160.353 26.757 −24.466 1.00 18.69 ATOM 2427 CZ TYR A 316 0.551 27.812−25.342 1.00 21.77 ATOM 2428 OH TYR A 316 1.002 27.557 −26.617 1.0023.75 ATOM 2429 CE2 TYR A 316 0.329 29.125 −24.932 1.00 22.39 ATOM 2430CD2 TYR A 316 −0.111 29.369 −23.639 1.00 21.90 ATOM 2431 C TYR A 316−0.037 29.730 −19.173 1.00 17.87 ATOM 2432 O TYR A 316 −0.968 30.517−19.266 1.00 17.06 ATOM 2433 N ASN A 317 0.689 29.555 −18.066 1.00 18.34ATOM 2434 CA ASN A 317 0.483 30.231 −16.766 1.00 19.27 ATOM 2435 CB ASNA 317 0.106 31.699 −16.921 1.00 20.01 ATOM 2436 CG ASN A 317 1.17132.489 −17.624 1.00 24.51 ATOM 2437 OD1 ASN A 317 2.363 32.384 −17.3051.00 29.46 ATOM 2438 ND2 ASN A 317 0.756 33.269 −18.603 1.00 29.08 ATOM2439 C ASN A 317 −0.506 29.551 −15.842 1.00 18.28 ATOM 2440 O ASN A 317−0.706 30.001 −14.719 1.00 19.05 ATOM 2441 N GLY A 318 −1.114 28.459−16.300 1.00 17.74 ATOM 2442 CA GLY A 318 −2.086 27.721 −15.475 1.0015.34 ATOM 2443 C GLY A 318 −3.458 28.356 −15.550 1.00 15.47 ATOM 2444 OGLY A 318 −3.700 29.390 −14.932 1.00 15.75 ATOM 2445 N ASN A 319 −4.36927.733 −16.306 1.00 13.16 ATOM 2446 CA ASN A 319 −5.672 28.305 −16.5571.00 12.74 ATOM 2447 CB ASN A 319 −5.693 28.883 −17.980 1.00 12.31 ATOM2448 CG ASN A 319 −4.676 29.979 −18.187 1.00 13.01 ATOM 2449 OD1 ASN A319 −4.832 31.117 −17.699 1.00 14.18 ATOM 2450 ND2 ASN A 319 −3.64029.665 −18.942 1.00 11.49 ATOM 2451 C ASN A 319 −6.799 27.271 −16.4421.00 12.27 ATOM 2452 O ASN A 319 −6.545 26.071 −16.456 1.00 12.28 ATOM2453 N PRO A 320 −8.054 27.732 −16.334 1.00 12.96 ATOM 2454 CA PRO A 320−9.113 26.759 −16.472 1.00 12.58 ATOM 2455 CB PRO A 320 −10.395 27.579−16.324 1.00 13.28 ATOM 2456 CG PRO A 320 −10.007 29.011 −16.183 1.0014.34 ATOM 2457 CD PRO A 320 −8.537 29.090 −15.991 1.00 12.66 ATOM 2458C PRO A 320 −9.101 26.090 −17.851 1.00 12.18 ATOM 2459 O PRO A 320−8.643 26.698 −18.820 1.00 11.99 ATOM 2460 N TRP A 321 −9.589 24.852−17.912 1.00 11.79 ATOM 2461 CA TRP A 321 −9.739 24.116 −19.154 1.0011.97 ATOM 2462 CB TRP A 321 −8.988 22.775 −19.063 1.00 11.15 ATOM 2463CG TRP A 321 −7.469 22.900 −19.200 1.00 12.16 ATOM 2464 CD1 TRP A 321−6.658 23.837 −18.627 1.00 13.28 ATOM 2465 NE1 TRP A 321 −5.347 23.636−19.016 1.00 13.24 ATOM 2466 CE2 TRP A 321 −5.290 22.538 −19.831 1.0013.51 ATOM 2467 CD2 TRP A 321 −6.617 22.054 −19.978 1.00 12.68 ATOM 2468CE3 TRP A 321 −6.846 20.938 −20.787 1.00 14.41 ATOM 2469 CZ3 TRP A 321−5.741 20.323 −21.428 1.00 13.49 ATOM 2470 CH2 TRP A 321 −4.436 20.819−21.250 1.00 13.43 ATOM 2471 CZ2 TRP A 321 −4.193 21.948 −20.479 1.0014.69 ATOM 2472 C TRP A 321 −11.202 23.797 −19.342 1.00 11.99 ATOM 2473O TRP A 321 −11.875 23.448 −18.388 1.00 11.51 ATOM 2474 N TYR A 322−11.696 23.896 −20.579 1.00 11.47 ATOM 2475 CA TYR A 322 −13.088 23.511−20.841 1.00 11.31 ATOM 2476 CB TYR A 322 −13.433 23.691 −22.322 1.0012.14 ATOM 2477 CG TYR A 322 −13.352 25.130 −22.793 1.00 12.81 ATOM 2478CD1 TYR A 322 −12.260 25.574 −23.509 1.00 11.43 ATOM 2479 CE1 TYR A 322−12.173 26.914 −23.965 1.00 12.91 ATOM 2480 CZ TYR A 322 −13.216 27.802−23.697 1.00 14.27 ATOM 2481 OH TYR A 322 −13.127 29.104 −24.146 1.0015.10 ATOM 2482 CE2 TYR A 322 −14.324 27.373 −22.982 1.00 13.40 ATOM2483 CD2 TYR A 322 −14.378 26.031 −22.522 1.00 11.58 ATOM 2484 C TYR A322 −13.367 22.082 −20.433 1.00 11.31 ATOM 2485 O TYR A 322 −14.38021.795 −19.771 1.00 11.08 ATOM 2486 N LEU A 323 −12.480 21.169 −20.8141.00 10.96 ATOM 2487 CA LEU A 323 −12.770 19.750 −20.561 1.00 11.04 ATOM2488 CB LEU A 323 −11.787 18.844 −21.315 1.00 11.26 ATOM 2489 CG LEU A323 −10.314 18.876 −20.903 1.00 10.53 ATOM 2490 CD1 LEU A 323 −10.07417.902 −19.745 1.00 14.57 ATOM 2491 CD2 LEU A 323 −9.474 18.437 −22.1121.00 13.19 ATOM 2492 C LEU A 323 −12.778 19.449 −19.048 1.00 11.68 ATOM2493 O LEU A 323 −13.444 18.510 −18.602 1.00 12.06 ATOM 2494 N ALA A 324−12.036 20.239 −18.268 1.00 9.86 ATOM 2495 CA ALA A 324 −11.969 20.017−16.812 1.00 10.09 ATOM 2496 CB ALA A 324 −10.746 20.767 −16.234 1.009.76 ATOM 2497 C ALA A 324 −13.272 20.518 −16.178 1.00 10.17 ATOM 2498 OALA A 324 −13.866 19.840 −15.325 1.00 10.29 ATOM 2499 N THR A 325−13.758 21.662 −16.665 1.00 9.84 ATOM 2500 CA THR A 325 −15.000 22.267−16.172 1.00 11.14 ATOM 2501 CB THR A 325 −15.102 23.765 −16.623 1.0012.15 ATOM 2502 OG1 THR A 325 −14.002 24.498 −16.063 1.00 13.16 ATOM2503 CG2 THR A 325 −16.402 24.411 −16.152 1.00 11.83 ATOM 2504 C THR A325 −16.218 21.413 −16.570 1.00 11.50 ATOM 2505 O THR A 325 −17.08621.126 −15.727 1.00 10.79 ATOM 2506 N PHE A 326 −16.234 20.925 −17.8161.00 10.79 ATOM 2507 CA PHE A 326 −17.272 19.959 −18.240 1.00 12.12 ATOM2508 CB PHE A 326 −17.194 19.652 −19.746 1.00 12.14 ATOM 2509 CG PHE A326 −17.518 20.851 −20.640 1.00 13.71 ATOM 2510 CD1 PHE A 326 −16.77721.077 −21.804 1.00 15.21 ATOM 2511 CE1 PHE A 326 −17.043 22.188 −22.6351.00 14.99 ATOM 2512 CZ PHE A 326 −18.072 23.066 −22.311 1.00 16.33 ATOM2513 CE2 PHE A 326 −18.851 22.832 −21.160 1.00 20.12 ATOM 2514 CD2 PHE A326 −18.561 21.717 −20.331 1.00 16.63 ATOM 2515 C PHE A 326 −17.21618.643 −17.464 1.00 11.56 ATOM 2516 O PHE A 326 −18.263 18.069 −17.1801.00 11.74 ATOM 2517 N ALA A 327 −16.014 18.174 −17.103 1.00 11.35 ATOM2518 CA ALA A 327 −15.889 16.909 −16.346 1.00 11.41 ATOM 2519 CB ALA A327 −14.397 16.538 −16.158 1.00 11.95 ATOM 2520 C ALA A 327 −16.61216.964 −14.965 1.00 12.04 ATOM 2521 O ALA A 327 −17.260 15.985 −14.5611.00 12.69 ATOM 2522 N ALA A 328 −16.505 18.097 −14.266 1.00 12.13 ATOM2523 CA ALA A 328 −17.207 18.293 −12.985 1.00 12.24 ATOM 2524 CB ALA A328 −16.871 19.662 −12.369 1.00 12.48 ATOM 2525 C ALA A 328 −18.70718.157 −13.177 1.00 12.90 ATOM 2526 O ALA A 328 −19.378 17.454 −12.4111.00 13.60 ATOM 2527 N ALA A 329 −19.239 18.814 −14.202 1.00 12.55 ATOM2528 CA ALA A 329 −20.669 18.682 −14.504 1.00 12.31 ATOM 2529 CB ALA A329 −21.027 19.551 −15.692 1.00 12.87 ATOM 2530 C ALA A 329 −21.03517.226 −14.788 1.00 12.71 ATOM 2531 O ALA A 329 −22.016 16.700 −14.2661.00 12.32 ATOM 2532 N GLU A 330 −20.231 16.572 −15.629 1.00 12.54 ATOM2533 CA GLU A 330 −20.500 15.187 −16.003 1.00 12.84 ATOM 2534 CB GLU A330 −19.519 14.718 −17.100 1.00 12.55 ATOM 2535 CG GLU A 330 −19.85013.303 −17.626 1.00 13.80 ATOM 2536 CD GLU A 330 −19.108 12.953 −18.9171.00 13.54 ATOM 2537 OE1 GLU A 330 −18.650 13.889 −19.604 1.00 12.29ATOM 2538 OE2 GLU A 330 −18.998 11.739 −19.209 1.00 14.52 ATOM 2539 CGLU A 330 −20.523 14.231 −14.809 1.00 12.94 ATOM 2540 O GLU A 330−21.400 13.346 −14.726 1.00 12.90 ATOM 2541 N GLN A 331 −19.598 14.402−13.866 1.00 12.03 ATOM 2542 CA GLN A 331 −19.589 13.502 −12.726 1.0012.38 ATOM 2543 CB GLN A 331 −18.415 13.795 −11.797 1.00 12.24 ATOM 2544CG GLN A 331 −18.357 12.759 −10.670 1.00 13.61 ATOM 2545 CD GLN A 331−17.327 13.072 −9.608 1.00 15.82 ATOM 2546 OE1 GLN A 331 −16.263 13.617−9.895 1.00 15.39 ATOM 2547 NE2 GLN A 331 −17.628 12.702 −8.372 1.0013.76 ATOM 2548 C GLN A 331 −20.912 13.643 −11.969 1.00 12.33 ATOM 2549O GLN A 331 −21.512 12.659 −11.556 1.00 12.45 ATOM 2550 N LEU A 332−21.377 14.873 −11.844 1.00 12.57 ATOM 2551 CA LEU A 332 −22.628 15.138−11.134 1.00 13.59 ATOM 2552 CB LEU A 332 −22.747 16.631 −10.868 1.0013.17 ATOM 2553 CG LEU A 332 −21.681 17.142 −9.867 1.00 16.56 ATOM 2554CD1 LEU A 332 −21.718 18.678 −9.801 1.00 18.10 ATOM 2555 CD2 LEU A 332−21.851 16.476 −8.492 1.00 19.47 ATOM 2556 C LEU A 332 −23.861 14.600−11.864 1.00 13.57 ATOM 2557 O LEU A 332 −24.770 14.053 −11.239 1.0013.24 ATOM 2558 N TYR A 333 −23.909 14.766 −13.179 1.00 13.90 ATOM 2559CA TYR A 333 −24.988 14.131 −13.972 1.00 14.37 ATOM 2560 CB TYR A 333−24.901 14.523 −15.468 1.00 14.48 ATOM 2561 CG TYR A 333 −25.056 16.001−15.721 1.00 13.91 ATOM 2562 CD1 TYR A 333 −26.086 16.738 −15.118 1.0014.64 ATOM 2563 CE1 TYR A 333 −26.208 18.117 −15.350 1.00 15.65 ATOM2564 CZ TYR A 333 −25.315 18.758 −16.196 1.00 16.47 ATOM 2565 OH TYR A333 −25.431 20.101 −16.442 1.00 17.22 ATOM 2566 CE2 TYR A 333 −24.31018.050 −16.836 1.00 16.59 ATOM 2567 CD2 TYR A 333 −24.192 16.669 −16.6011.00 11.27 ATOM 2568 C TYR A 333 −25.022 12.613 −13.843 1.00 15.03 ATOM2569 O TYR A 333 −26.108 12.012 −13.824 1.00 14.78 ATOM 2570 N ASP A 334−23.836 11.998 −13.807 1.00 14.06 ATOM 2571 CA ASP A 334 −23.714 10.555−13.602 1.00 14.83 ATOM 2572 CB ASP A 334 −22.239 10.114 −13.714 1.0013.83 ATOM 2573 CG ASP A 334 −21.708 10.149 −15.136 1.00 15.84 ATOM 2574OD1 ASP A 334 −22.495 10.373 −16.081 1.00 13.95 ATOM 2575 OD2 ASP A 334−20.470 9.943 −15.313 1.00 15.50 ATOM 2576 C ASP A 334 −24.254 10.163−12.224 1.00 15.21 ATOM 2577 O ASP A 334 −24.941 9.132 −12.080 1.0015.93 ATOM 2578 N ALA A 335 −23.933 10.969 −11.213 1.00 15.09 ATOM 2579CA ALA A 335 −24.454 10.735 −9.855 1.00 16.00 ATOM 2580 CB ALA A 335−23.809 11.719 −8.864 1.00 15.13 ATOM 2581 C ALA A 335 −25.980 10.823−9.803 1.00 16.10 ATOM 2582 O ALA A 335 −26.643 9.916 −9.245 1.00 16.77ATOM 2583 N ILE A 336 −26.530 11.879 −10.398 1.00 16.35 ATOM 2584 CA ILEA 336 −27.987 12.087 −10.470 1.00 18.39 ATOM 2585 CB ILE A 336 −28.33213.422 −11.162 1.00 18.85 ATOM 2586 CG1 ILE A 336 −27.891 14.596 −10.2791.00 19.14 ATOM 2587 CD1 ILE A 336 −27.879 15.904 −10.986 1.00 22.50ATOM 2588 CG2 ILE A 336 −29.839 13.539 −11.506 1.00 20.14 ATOM 2589 CILE A 336 −28.681 10.902 −11.156 1.00 18.83 ATOM 2590 O ILE A 336−29.707 10.404 −10.675 1.00 18.06 ATOM 2591 N TYR A 337 −28.102 10.443−12.267 1.00 18.50 ATOM 2592 CA TYR A 337 −28.642 9.287 −12.970 1.0018.99 ATOM 2593 CB TYR A 337 −27.753 8.908 −14.169 1.00 19.80 ATOM 2594CG TYR A 337 −28.328 7.737 −14.954 1.00 20.76 ATOM 2595 CD1 TYR A 337−27.988 6.429 −14.620 1.00 20.95 ATOM 2596 CE1 TYR A 337 −28.511 5.345−15.322 1.00 22.94 ATOM 2597 CZ TYR A 337 −29.382 5.559 −16.356 1.0022.19 ATOM 2598 OH TYR A 337 −29.877 4.447 −17.018 1.00 24.87 ATOM 2599CE2 TYR A 337 −29.752 6.845 −16.721 1.00 22.31 ATOM 2600 CD2 TYR A 337−29.220 7.942 −16.009 1.00 21.58 ATOM 2601 C TYR A 337 −28.839 8.083−12.057 1.00 18.60 ATOM 2602 O TYR A 337 −29.918 7.476 −12.041 1.0018.61 ATOM 2603 N VAL A 338 −27.802 7.737 −11.297 1.00 18.67 ATOM 2604CA VAL A 338 −27.837 6.573 −10.406 1.00 18.90 ATOM 2605 CB VAL A 338−26.424 6.195 −9.919 1.00 18.99 ATOM 2606 CG1 VAL A 338 −26.462 5.121−8.820 1.00 19.71 ATOM 2607 CG2 VAL A 338 −25.600 5.698 −11.111 1.0018.75 ATOM 2608 C VAL A 338 −28.810 6.788 −9.234 1.00 19.41 ATOM 2609 OVAL A 338 −29.565 5.871 −8.869 1.00 19.45 ATOM 2610 N TRP A 339 −28.7977.987 −8.654 1.00 19.81 ATOM 2611 CA TRP A 339 −29.743 8.290 −7.559 1.0020.46 ATOM 2612 CB TRP A 339 −29.514 9.705 −7.029 1.00 20.35 ATOM 2613CG TRP A 339 −28.222 9.830 −6.329 1.00 18.64 ATOM 2614 CD1 TRP A 339−27.540 8.846 −5.676 1.00 16.51 ATOM 2615 NE1 TRP A 339 −26.391 9.359−5.126 1.00 17.81 ATOM 2616 CE2 TRP A 339 −26.312 10.693 −5.423 1.0017.12 ATOM 2617 CD2 TRP A 339 −27.452 11.025 −6.183 1.00 17.64 ATOM 2618CE3 TRP A 339 −27.624 12.343 −6.614 1.00 17.59 ATOM 2619 CZ3 TRP A 339−26.637 13.283 −6.284 1.00 19.24 ATOM 2620 CH2 TRP A 339 −25.510 12.912−5.520 1.00 18.24 ATOM 2621 CZ2 TRP A 339 −25.320 11.626 −5.103 1.0018.45 ATOM 2622 C TRP A 339 −31.201 8.108 −7.997 1.00 21.83 ATOM 2623 OTRP A 339 −31.981 7.478 −7.274 1.00 22.01 ATOM 2624 N LYS A 340 −31.5498.646 −9.168 1.00 22.85 ATOM 2625 CA LYS A 340 −32.904 8.541 −9.721 1.0025.61 ATOM 2626 CB LYS A 340 −33.066 9.411 −10.967 1.00 25.52 ATOM 2627CG LYS A 340 −33.174 10.905 −10.689 1.00 28.19 ATOM 2628 CD LYS A 340−33.227 11.692 −11.991 1.00 34.04 ATOM 2629 CE LYS A 340 −33.966 13.011−11.805 1.00 38.04 ATOM 2630 NZ LYS A 340 −33.868 13.876 −13.017 1.0041.83 ATOM 2631 C LYS A 340 −33.276 7.108 −10.062 1.00 27.14 ATOM 2632 OLYS A 340 −34.413 6.686 −9.830 1.00 27.56 ATOM 2633 N LYS A 341 −32.3176.358 −10.604 1.00 28.13 ATOM 2634 CA LYS A 341 −32.552 4.975 −11.0181.00 30.18 ATOM 2635 CB LYS A 341 −31.358 4.428 −11.800 1.00 29.83 ATOM2636 CG LYS A 341 −31.688 3.173 −12.624 1.00 33.04 ATOM 2637 CD LYS A341 −30.472 2.624 −13.395 1.00 33.62 ATOM 2638 CE LYS A 341 −29.6521.592 −12.588 1.00 38.22 ATOM 2639 NZ LYS A 341 −28.691 2.188 −11.5731.00 40.88 ATOM 2640 C LYS A 341 −32.816 4.081 −9.817 1.00 30.19 ATOM2641 O LYS A 341 −33.744 3.260 −9.837 1.00 30.15 ATOM 2642 N THR A 342−31.999 4.246 −8.777 1.00 29.52 ATOM 2643 CA THR A 342 −32.074 3.400−7.595 1.00 29.75 ATOM 2644 CB THR A 342 −30.687 3.221 −6.916 1.00 29.68ATOM 2645 OG1 THR A 342 −30.254 4.458 −6.333 1.00 32.01 ATOM 2646 CG2THR A 342 −29.628 2.735 −7.929 1.00 31.40 ATOM 2647 C THR A 342 −33.1293.901 −6.596 1.00 29.01 ATOM 2648 O THR A 342 −33.572 3.148 −5.734 1.0029.92 ATOM 2649 N GLY A 343 −33.534 5.158 −6.732 1.00 28.20 ATOM 2650 CAGLY A 343 −34.537 5.782 −5.862 1.00 28.30 ATOM 2651 C GLY A 343 −34.0686.045 −4.438 1.00 27.91 ATOM 2652 O GLY A 343 −34.887 6.133 −3.519 1.0028.16 ATOM 2653 N SER A 344 −32.760 6.226 −4.260 1.00 27.22 ATOM 2654 CASER A 344 −32.142 6.306 −2.939 1.00 26.60 ATOM 2655 CB SER A 344 −31.8704.880 −2.462 1.00 27.28 ATOM 2656 OG SER A 344 −31.354 4.855 −1.161 1.0029.50 ATOM 2657 C SER A 344 −30.823 7.107 −2.979 1.00 26.02 ATOM 2658 OSER A 344 −30.068 6.992 −3.944 1.00 25.80 ATOM 2659 N ILE A 345 −30.5577.900 −1.936 1.00 24.20 ATOM 2660 CA ILE A 345 −29.295 8.641 −1.770 1.0023.04 ATOM 2661 CB ILE A 345 −29.477 10.171 −1.954 1.00 23.03 ATOM 2662CG1 ILE A 345 −30.021 10.474 −3.340 1.00 22.44 ATOM 2663 CD1 ILE A 345−30.399 11.918 −3.599 1.00 23.24 ATOM 2664 CG2 ILE A 345 −28.138 10.918−1.670 1.00 22.00 ATOM 2665 C ILE A 345 −28.726 8.415 −0.378 1.00 23.13ATOM 2666 O ILE A 345 −29.392 8.684 0.623 1.00 23.57 ATOM 2667 N THR A346 −27.490 7.943 −0.307 1.00 22.23 ATOM 2668 CA THR A 346 −26.820 7.7650.963 1.00 23.25 ATOM 2669 CB THR A 346 −26.246 6.338 1.101 1.00 23.78ATOM 2670 OG1 THR A 346 −27.327 5.396 1.020 1.00 27.42 ATOM 2671 CG2 THRA 346 −25.507 6.129 2.443 1.00 24.74 ATOM 2672 C THR A 346 −25.753 8.8491.138 1.00 23.03 ATOM 2673 O THR A 346 −24.916 9.067 0.260 1.00 23.47ATOM 2674 N VAL A 347 −25.848 9.561 2.255 1.00 21.33 ATOM 2675 CA VAL A347 −24.845 10.537 2.674 1.00 20.11 ATOM 2676 CB VAL A 347 −25.52211.844 3.212 1.00 19.06 ATOM 2677 CG1 VAL A 347 −24.489 12.834 3.7001.00 19.66 ATOM 2678 CG2 VAL A 347 −26.418 12.465 2.137 1.00 20.32 ATOM2679 C VAL A 347 −24.066 9.865 3.785 1.00 20.14 ATOM 2680 O VAL A 347−24.667 9.340 4.728 1.00 19.79 ATOM 2681 N THR A 348 −22.734 9.878 3.6711.00 19.85 ATOM 2682 CA THR A 348 −21.851 9.274 4.660 1.00 19.92 ATOM2683 CB THR A 348 −20.965 8.185 4.018 1.00 19.82 ATOM 2684 OG1 THR A 348−19.921 8.815 3.277 1.00 20.35 ATOM 2685 CG2 THR A 348 −21.785 7.2783.092 1.00 21.67 ATOM 2686 C THR A 348 −20.964 10.354 5.256 1.00 19.60ATOM 2687 O THR A 348 −20.961 11.484 4.760 1.00 19.51 ATOM 2688 N ALA A349 −20.191 10.006 6.292 1.00 19.18 ATOM 2689 CA ALA A 349 −19.24310.932 6.885 1.00 20.02 ATOM 2690 CB ALA A 349 −18.494 10.275 8.044 1.0020.61 ATOM 2691 C ALA A 349 −18.240 11.466 5.842 1.00 19.79 ATOM 2692 OALA A 349 −17.756 12.601 5.947 1.00 20.15 ATOM 2693 N THR A 350 −17.90610.619 4.873 1.00 18.87 ATOM 2694 CA THR A 350 −16.911 10.971 3.850 1.0018.30 ATOM 2695 CB THR A 350 −16.435 9.717 3.093 1.00 18.67 ATOM 2696OG1 THR A 350 −15.780 8.850 4.027 1.00 19.82 ATOM 2697 CG2 THR A 350−15.426 10.097 1.974 1.00 17.73 ATOM 2698 C THR A 350 −17.463 12.0032.871 1.00 17.30 ATOM 2699 O THR A 350 −16.747 12.930 2.487 1.00 17.99ATOM 2700 N SER A 351 −18.716 11.847 2.467 1.00 16.16 ATOM 2701 CA SER A351 −19.316 12.803 1.517 1.00 15.78 ATOM 2702 CB SER A 351 −20.21412.076 0.512 1.00 15.85 ATOM 2703 OG SER A 351 −21.280 11.412 1.156 1.0017.14 ATOM 2704 C SER A 351 −20.087 13.941 2.193 1.00 15.63 ATOM 2705 OSER A 351 −20.736 14.743 1.524 1.00 13.62 ATOM 2706 N LEU A 352 −20.04814.006 3.527 1.00 15.09 ATOM 2707 CA LEU A 352 −20.901 14.985 4.212 1.0016.29 ATOM 2708 CB LEU A 352 −20.759 14.851 5.736 1.00 16.64 ATOM 2709CG LEU A 352 −21.713 15.734 6.570 1.00 17.22 ATOM 2710 CD1 LEU A 352−23.138 15.281 6.418 1.00 19.06 ATOM 2711 CD2 LEU A 352 −21.263 15.6368.032 1.00 20.02 ATOM 2712 C LEU A 352 −20.592 16.427 3.787 1.00 16.09ATOM 2713 O LEU A 352 −21.499 17.219 3.601 1.00 16.35 ATOM 2714 N ALA A353 −19.311 16.763 3.643 1.00 16.19 ATOM 2715 CA ALA A 353 −18.93318.148 3.354 1.00 15.74 ATOM 2716 CB ALA A 353 −17.460 18.314 3.424 1.0016.30 ATOM 2717 C ALA A 353 −19.459 18.544 1.972 1.00 16.11 ATOM 2718 OALA A 353 −19.957 19.668 1.781 1.00 15.44 ATOM 2719 N PHE A 354 −19.36717.607 1.021 1.00 16.10 ATOM 2720 CA PHE A 354 −19.885 17.849 −0.3251.00 15.25 ATOM 2721 CB PHE A 354 −19.718 16.618 −1.220 1.00 16.59 ATOM2722 CG PHE A 354 −20.497 16.707 −2.500 1.00 15.45 ATOM 2723 CD1 PHE A354 −19.959 17.375 −3.603 1.00 16.95 ATOM 2724 CE1 PHE A 354 −20.66417.489 −4.793 1.00 17.06 ATOM 2725 CZ PHE A 354 −21.956 16.953 −4.8881.00 16.75 ATOM 2726 CE2 PHE A 354 −22.517 16.276 −3.791 1.00 16.91 ATOM2727 CD2 PHE A 354 −21.778 16.160 −2.594 1.00 17.48 ATOM 2728 C PHE A354 −21.374 18.188 −0.226 1.00 15.56 ATOM 2729 O PHE A 354 −21.81519.183 −0.797 1.00 15.08 ATOM 2730 N PHE A 355 −22.140 17.347 0.474 1.0014.54 ATOM 2731 CA PHE A 355 −23.588 17.544 0.517 1.00 15.29 ATOM 2732CB PHE A 355 −24.295 16.319 1.078 1.00 15.61 ATOM 2733 CG PHE A 355−24.386 15.176 0.112 1.00 15.51 ATOM 2734 CD1 PHE A 355 −25.306 15.205−0.945 1.00 14.91 ATOM 2735 CE1 PHE A 355 −25.404 14.131 −1.832 1.0016.52 ATOM 2736 CZ PHE A 355 −24.567 13.033 −1.676 1.00 16.12 ATOM 2737CE2 PHE A 355 −23.648 12.994 −0.628 1.00 15.62 ATOM 2738 CD2 PHE A 355−23.562 14.071 0.255 1.00 13.62 ATOM 2739 C PHE A 355 −23.988 18.7891.303 1.00 15.19 ATOM 2740 O PHE A 355 −24.920 19.477 0.902 1.00 15.69ATOM 2741 N GLN A 356 −23.283 19.084 2.398 1.00 15.78 ATOM 2742 CA GLN A356 −23.679 20.216 3.257 1.00 16.16 ATOM 2743 CB GLN A 356 −22.98720.165 4.627 1.00 16.84 ATOM 2744 CG GLN A 356 −23.564 19.115 5.579 1.0017.13 ATOM 2745 CD GLN A 356 −22.907 19.170 6.973 1.00 18.06 ATOM 2746OE1 GLN A 356 −21.808 19.707 7.138 1.00 20.29 ATOM 2747 NE2 GLN A 356−23.569 18.589 7.965 1.00 20.95 ATOM 2748 C GLN A 356 −23.444 21.5462.584 1.00 16.08 ATOM 2749 O GLN A 356 −24.142 22.507 2.868 1.00 15.59ATOM 2750 N GLU A 357 −22.482 21.599 1.661 1.00 16.07 ATOM 2751 CA GLU A357 −22.261 22.808 0.884 1.00 16.41 ATOM 2752 CB GLU A 357 −20.99422.683 0.005 1.00 16.33 ATOM 2753 CG GLU A 357 −20.671 23.942 −0.7701.00 15.64 ATOM 2754 CD GLU A 357 −19.326 23.894 −1.516 1.00 17.67 ATOM2755 OE1 GLU A 357 −18.931 24.947 −2.066 1.00 19.03 ATOM 2756 OE2 GLU A357 −18.685 22.822 −1.575 1.00 14.75 ATOM 2757 C GLU A 357 −23.49223.105 0.019 1.00 15.84 ATOM 2758 O GLU A 357 −23.786 24.237 −0.224 1.0018.07 ATOM 2759 N LEU A 358 −24.213 22.084 −0.420 1.00 14.72 ATOM 2760CA LEU A 358 −25.364 22.251 −1.310 1.00 15.75 ATOM 2761 CB LEU A 358−25.368 21.147 −2.369 1.00 16.49 ATOM 2762 CG LEU A 358 −24.057 21.100−3.168 1.00 16.81 ATOM 2763 CD1 LEU A 358 −24.087 19.977 −4.182 1.0019.73 ATOM 2764 CD2 LEU A 358 −23.775 22.465 −3.846 1.00 19.34 ATOM 2765C LEU A 358 −26.708 22.251 −0.582 1.00 15.36 ATOM 2766 O LEU A 358−27.656 22.911 −1.028 1.00 14.78 ATOM 2767 N VAL A 359 −26.786 21.5110.520 1.00 15.34 ATOM 2768 CA VAL A 359 −28.001 21.404 1.321 1.00 15.00ATOM 2769 CB VAL A 359 −28.691 20.006 1.154 1.00 15.90 ATOM 2770 CG1 VALA 359 −29.999 19.917 1.962 1.00 15.16 ATOM 2771 CG2 VAL A 359 −28.96719.685 −0.348 1.00 16.26 ATOM 2772 C VAL A 359 −27.531 21.624 2.775 1.0014.72 ATOM 2773 O VAL A 359 −27.192 20.653 3.500 1.00 14.22 ATOM 2774 NPRO A 360 −27.467 22.893 3.193 1.00 14.98 ATOM 2775 CA PRO A 360 −26.93723.179 4.539 1.00 15.45 ATOM 2776 CB PRO A 360 −27.150 24.700 4.700 1.0015.45 ATOM 2777 CG PRO A 360 −27.188 25.219 3.274 1.00 16.15 ATOM 2778CD PRO A 360 −27.854 24.127 2.471 1.00 14.54 ATOM 2779 C PRO A 360−27.692 22.385 5.611 1.00 15.31 ATOM 2780 O PRO A 360 −28.918 22.2625.555 1.00 15.07 ATOM 2781 N GLY A 361 −26.936 21.842 6.560 1.00 15.94ATOM 2782 CA GLY A 361 −27.512 21.143 7.709 1.00 16.53 ATOM 2783 C GLY A361 −27.870 19.680 7.488 1.00 17.03 ATOM 2784 O GLY A 361 −28.268 18.9978.429 1.00 17.68 ATOM 2785 N VAL A 362 −27.762 19.176 6.261 1.00 16.13ATOM 2786 CA VAL A 362 −28.163 17.769 6.037 1.00 16.72 ATOM 2787 CB VALA 362 −28.217 17.416 4.525 1.00 16.47 ATOM 2788 CG1 VAL A 362 −26.80817.311 3.947 1.00 16.75 ATOM 2789 CG2 VAL A 362 −29.054 16.142 4.2801.00 17.21 ATOM 2790 C VAL A 362 −27.208 16.849 6.811 1.00 17.24 ATOM2791 O VAL A 362 −26.044 17.187 7.006 1.00 17.07 ATOM 2792 N THR A 363−27.695 15.703 7.274 1.00 18.15 ATOM 2793 CA THR A 363 −26.821 14.7898.025 1.00 19.86 ATOM 2794 CB THR A 363 −27.388 14.459 9.405 1.00 20.59ATOM 2795 OG1 THR A 363 −28.634 13.776 9.217 1.00 22.85 ATOM 2796 CG2THR A 363 −27.610 15.742 10.182 1.00 22.71 ATOM 2797 C THR A 363 −26.66013.476 7.310 1.00 19.18 ATOM 2798 O THR A 363 −27.398 13.184 6.371 1.0018.87 ATOM 2799 N ALA A 364 −25.697 12.679 7.769 1.00 19.67 ATOM 2800 CAALA A 364 −25.495 11.342 7.222 1.00 20.19 ATOM 2801 CB ALA A 364 −24.36110.637 7.930 1.00 20.38 ATOM 2802 C ALA A 364 −26.783 10.541 7.343 1.0021.17 ATOM 2803 O ALA A 364 −27.551 10.720 8.293 1.00 21.92 ATOM 2804 NGLY A 365 −27.041 9.687 6.360 1.00 21.67 ATOM 2805 CA GLY A 365 −28.2078.823 6.371 1.00 22.35 ATOM 2806 C GLY A 365 −28.584 8.415 4.968 1.0023.66 ATOM 2807 O GLY A 365 −27.924 8.804 3.991 1.00 23.66 ATOM 2808 NTHR A 366 −29.639 7.615 4.862 1.00 23.70 ATOM 2809 CA THR A 366 −30.1487.183 3.582 1.00 24.56 ATOM 2810 CB THR A 366 −30.188 5.644 3.491 1.0025.67 ATOM 2811 OG1 THR A 366 −28.849 5.143 3.649 1.00 27.09 ATOM 2812CG2 THR A 366 −30.715 5.216 2.159 1.00 25.47 ATOM 2813 C THR A 366−31.520 7.769 3.344 1.00 25.15 ATOM 2814 O THR A 366 −32.427 7.612 4.1771.00 25.01 ATOM 2815 N TYR A 367 −31.668 8.447 2.210 1.00 24.20 ATOM2816 CA TYR A 367 −32.900 9.146 1.883 1.00 24.30 ATOM 2817 CB TYR A 367−32.616 10.648 1.701 1.00 23.51 ATOM 2818 CG TYR A 367 −31.924 11.2382.907 1.00 22.68 ATOM 2819 CD1 TYR A 367 −32.639 11.506 4.078 1.00 21.99ATOM 2820 CE1 TYR A 367 −32.012 12.019 5.199 1.00 20.27 ATOM 2821 CZ TYRA 367 −30.650 12.263 5.176 1.00 22.67 ATOM 2822 OH TYR A 367 −30.03612.789 6.287 1.00 21.24 ATOM 2823 CE2 TYR A 367 −29.897 11.994 4.0231.00 20.71 ATOM 2824 CD2 TYR A 367 −30.541 11.479 2.904 1.00 20.37 ATOM2825 C TYR A 367 −33.531 8.542 0.641 1.00 25.39 ATOM 2826 O TYR A 367−32.900 8.456 −0.415 1.00 24.86 ATOM 2827 N SER A 368 −34.782 8.1090.758 1.00 25.82 ATOM 2828 CA SER A 368 −35.416 7.454 −0.374 1.00 27.29ATOM 2829 CB SER A 368 −36.339 6.332 0.107 1.00 27.90 ATOM 2830 OG SER A368 −37.519 6.895 0.634 1.00 30.69 ATOM 2831 C SER A 368 −36.171 8.466−1.218 1.00 27.68 ATOM 2832 O SER A 368 −36.361 9.612 −0.805 1.00 26.96ATOM 2833 N SER A 369 −36.629 8.025 −2.388 1.00 28.98 ATOM 2834 CA SER A369 −37.260 8.908 −3.367 1.00 30.52 ATOM 2835 CB SER A 369 −37.520 8.167−4.681 1.00 30.90 ATOM 2836 OG SER A 369 −38.269 6.983 −4.452 1.00 32.57ATOM 2837 C SER A 369 −38.536 9.583 −2.871 1.00 31.50 ATOM 2838 O SER A369 −38.954 10.572 −3.442 1.00 32.33 ATOM 2839 N SER A 370 −39.150 9.067−1.809 1.00 32.18 ATOM 2840 CA SER A 370 −40.365 9.712 −1.279 1.00 32.96ATOM 2841 CB SER A 370 −41.313 8.692 −0.624 1.00 33.18 ATOM 2842 OG SERA 370 −40.610 7.847 0.273 1.00 34.18 ATOM 2843 C SER A 370 −40.04910.864 −0.323 1.00 32.37 ATOM 2844 O SER A 370 −40.901 11.729 −0.0781.00 33.26 ATOM 2845 N SER A 371 −38.825 10.893 0.197 1.00 31.13 ATOM2846 CA SER A 371 −38.443 11.911 1.174 1.00 30.04 ATOM 2847 CB SER A 371−37.180 11.487 1.912 1.00 29.83 ATOM 2848 OG SER A 371 −36.046 11.7141.100 1.00 30.43 ATOM 2849 C SER A 371 −38.247 13.295 0.553 1.00 29.13ATOM 2850 O SER A 371 −37.795 13.424 −0.589 1.00 28.84 ATOM 2851 N SER A372 −38.571 14.340 1.312 1.00 27.84 ATOM 2852 CA SER A 372 −38.30015.689 0.845 1.00 27.18 ATOM 2853 CB SER A 372 −38.896 16.737 1.789 1.0027.36 ATOM 2854 OG SER A 372 −38.331 16.609 3.080 1.00 28.50 ATOM 2855 CSER A 372 −36.783 15.902 0.680 1.00 25.79 ATOM 2856 O SER A 372 −36.35816.690 −0.173 1.00 26.29 ATOM 2857 N THR A 373 −35.979 15.193 1.479 1.0024.06 ATOM 2858 CA THR A 373 −34.517 15.337 1.448 1.00 22.51 ATOM 2859CB THR A 373 −33.833 14.501 2.545 1.00 22.26 ATOM 2860 OG1 THR A 373−34.543 14.636 3.788 1.00 23.04 ATOM 2861 CG2 THR A 373 −32.370 14.9262.734 1.00 21.06 ATOM 2862 C THR A 373 −33.984 14.906 0.076 1.00 21.61ATOM 2863 O THR A 373 −33.134 15.578 −0.513 1.00 19.74 ATOM 2864 N PHE A374 −34.493 13.777 −0.413 1.00 21.13 ATOM 2865 CA PHE A 374 −34.13713.280 −1.749 1.00 22.10 ATOM 2866 CB PHE A 374 −34.950 12.023 −2.0511.00 22.12 ATOM 2867 CG PHE A 374 −34.624 11.380 −3.366 1.00 22.90 ATOM2868 CD1 PHE A 374 −33.677 10.368 −3.432 1.00 23.83 ATOM 2869 CE1 PHE A374 −33.381 9.749 −4.649 1.00 22.56 ATOM 2870 CZ PHE A 374 −34.04110.162 −5.802 1.00 23.24 ATOM 2871 CE2 PHE A 374 −34.985 11.161 −5.7521.00 23.36 ATOM 2872 CD2 PHE A 374 −35.280 11.769 −4.523 1.00 23.73 ATOM2873 C PHE A 374 −34.343 14.349 −2.818 1.00 22.24 ATOM 2874 O PHE A 374−33.413 14.681 −3.548 1.00 22.63 ATOM 2875 N THR A 375 −35.549 14.923−2.880 1.00 22.72 ATOM 2876 CA THR A 375 −35.890 15.963 −3.852 1.0023.34 ATOM 2877 CB THR A 375 −37.364 16.398 −3.683 1.00 23.63 ATOM 2878OG1 THR A 375 −38.193 15.244 −3.809 1.00 27.83 ATOM 2879 CG2 THR A 375−37.768 17.413 −4.749 1.00 27.05 ATOM 2880 C THR A 375 −35.003 17.203−3.746 1.00 22.66 ATOM 2881 O THR A 375 −34.603 17.766 −4.756 1.00 21.43ATOM 2882 N ASN A 376 −34.744 17.632 −2.508 1.00 20.80 ATOM 2883 CA ASNA 376 −33.880 18.766 −2.207 1.00 21.33 ATOM 2884 CB ASN A 376 −33.85618.975 −0.688 1.00 21.98 ATOM 2885 CG ASN A 376 −33.343 20.354 −0.2781.00 27.01 ATOM 2886 OD1 ASN A 376 −32.582 21.011 −1.004 1.00 31.72 ATOM2887 ND2 ASN A 376 −33.748 20.793 0.913 1.00 30.14 ATOM 2888 C ASN A 376−32.465 18.527 −2.733 1.00 19.80 ATOM 2889 O ASN A 376 −31.898 19.389−3.415 1.00 19.75 ATOM 2890 N ILE A 377 −31.915 17.354 −2.431 1.00 19.00ATOM 2891 CA ILE A 377 −30.586 16.983 −2.916 1.00 18.66 ATOM 2892 CB ILEA 377 −30.081 15.651 −2.319 1.00 18.56 ATOM 2893 CG1 ILE A 377 −29.83415.813 −0.800 1.00 18.42 ATOM 2894 CD1 ILE A 377 −29.634 14.481 −0.0281.00 19.35 ATOM 2895 CG2 ILE A 377 −28.787 15.233 −3.025 1.00 18.07 ATOM2896 C ILE A 377 −30.546 16.964 −4.451 1.00 18.87 ATOM 2897 O ILE A 377−29.655 17.575 −5.058 1.00 18.47 ATOM 2898 N ILE A 378 −31.513 16.293−5.068 1.00 18.45 ATOM 2899 CA ILE A 378 −31.556 16.216 −6.539 1.0019.78 ATOM 2900 CB ILE A 378 −32.738 15.359 −7.085 1.00 20.45 ATOM 2901CG1 ILE A 378 −32.593 13.891 −6.650 1.00 22.32 ATOM 2902 CD1 ILE A 378−31.414 13.145 −7.270 1.00 24.68 ATOM 2903 CG2 ILE A 378 −32.829 15.472−8.646 1.00 21.27 ATOM 2904 C ILE A 378 −31.561 17.588 −7.177 1.00 19.82ATOM 2905 O ILE A 378 −30.760 17.849 −8.101 1.00 19.62 ATOM 2906 N ASN A379 −32.441 18.470 −6.689 1.00 18.68 ATOM 2907 CA ASN A 379 −32.53119.820 −7.224 1.00 19.13 ATOM 2908 CB ASN A 379 −33.738 20.578 −6.6581.00 20.08 ATOM 2909 CG ASN A 379 −35.066 19.956 −7.087 1.00 25.11 ATOM2910 OD1 ASN A 379 −35.121 19.173 −8.044 1.00 29.37 ATOM 2911 ND2 ASN A379 −36.144 20.289 −6.369 1.00 29.08 ATOM 2912 C ASN A 379 −31.24120.604 −7.040 1.00 17.96 ATOM 2913 O ASN A 379 −30.774 21.273 −7.9811.00 17.85 ATOM 2914 N ALA A 380 −30.662 20.497 −5.841 1.00 15.93 ATOM2915 CA ALA A 380 −29.458 21.241 −5.509 1.00 16.08 ATOM 2916 CB ALA A380 −29.120 21.061 −4.033 1.00 16.14 ATOM 2917 C ALA A 380 −28.29920.783 −6.389 1.00 15.64 ATOM 2918 O ALA A 380 −27.566 21.607 −6.9381.00 16.56 ATOM 2919 N VAL A 381 −28.153 19.471 −6.519 1.00 15.22 ATOM2920 CA VAL A 381 −27.039 18.912 −7.302 1.00 15.77 ATOM 2921 CB VAL A381 −26.823 17.403 −6.999 1.00 15.61 ATOM 2922 CG1 VAL A 381 −25.74716.777 −7.940 1.00 14.83 ATOM 2923 CG2 VAL A 381 −26.386 17.234 −5.5511.00 14.84 ATOM 2924 C VAL A 381 −27.243 19.211 −8.794 1.00 16.08 ATOM2925 O VAL A 381 −26.281 19.508 −9.505 1.00 16.62 ATOM 2926 N SER A 382−28.482 19.112 −9.278 1.00 16.66 ATOM 2927 CA SER A 382 −28.772 19.453−10.690 1.00 18.67 ATOM 2928 CB SER A 382 −30.246 19.212 −11.043 1.0018.26 ATOM 2929 OG SER A 382 −30.538 17.855 −10.893 1.00 24.56 ATOM 2930C SER A 382 −28.434 20.894 −11.005 1.00 18.10 ATOM 2931 O SER A 382−27.815 21.183 −12.027 1.00 18.05 ATOM 2932 N THR A 383 −28.853 21.810−10.132 1.00 17.66 ATOM 2933 CA THR A 383 −28.521 23.216 −10.298 1.0017.72 ATOM 2934 CB THR A 383 −29.199 24.063 −9.180 1.00 18.48 ATOM 2935OG1 THR A 383 −30.606 23.985 −9.373 1.00 19.72 ATOM 2936 CG2 THR A 383−28.771 25.550 −9.227 1.00 19.59 ATOM 2937 C THR A 383 −27.017 23.470−10.314 1.00 17.09 ATOM 2938 O THR A 383 −26.524 24.286 −11.109 1.0017.00 ATOM 2939 N TYR A 384 −26.299 22.774 −9.435 1.00 15.62 ATOM 2940CA TYR A 384 −24.858 22.925 −9.312 1.00 15.37 ATOM 2941 CB TYR A 384−24.397 22.164 −8.068 1.00 15.00 ATOM 2942 CG TYR A 384 −22.958 22.345−7.630 1.00 15.08 ATOM 2943 CD1 TYR A 384 −22.361 23.601 −7.578 1.0015.83 ATOM 2944 CE1 TYR A 384 −21.049 23.752 −7.131 1.00 16.33 ATOM 2945CZ TYR A 384 −20.321 22.623 −6.737 1.00 16.13 ATOM 2946 OH TYR A 384−19.018 22.738 −6.302 1.00 16.22 ATOM 2947 CE2 TYR A 384 −20.890 21.386−6.778 1.00 13.72 ATOM 2948 CD2 TYR A 384 −22.203 21.242 −7.232 1.0014.72 ATOM 2949 C TYR A 384 −24.186 22.396 −10.590 1.00 15.13 ATOM 2950O TYR A 384 −23.319 23.065 −11.173 1.00 14.43 ATOM 2951 N ALA A 385−24.605 21.213 −11.033 1.00 15.02 ATOM 2952 CA ALA A 385 −24.045 20.639−12.263 1.00 15.60 ATOM 2953 CB ALA A 385 −24.634 19.295 −12.503 1.0015.85 ATOM 2954 C ALA A 385 −24.249 21.564 −13.477 1.00 16.15 ATOM 2955O ALA A 385 −23.292 21.857 −14.211 1.00 15.49 ATOM 2956 N ASP A 386−25.483 22.055 −13.660 1.00 15.57 ATOM 2957 CA ASP A 386 −25.782 23.058−14.694 1.00 16.09 ATOM 2958 CB ASP A 386 −27.279 23.433 −14.687 1.0015.88 ATOM 2959 CG ASP A 386 −28.158 22.379 −15.349 1.00 18.85 ATOM 2960OD1 ASP A 386 −27.672 21.307 −15.766 1.00 18.94 ATOM 2961 OD2 ASP A 386−29.365 22.624 −15.461 1.00 23.88 ATOM 2962 C ASP A 386 −24.938 24.322−14.526 1.00 15.81 ATOM 2963 O ASP A 386 −24.594 24.998 −15.501 1.0016.29 ATOM 2964 N GLY A 387 −24.591 24.640 −13.290 1.00 15.76 ATOM 2965CA GLY A 387 −23.735 25.787 −13.038 1.00 14.35 ATOM 2966 C GLY A 387−22.354 25.654 −13.663 1.00 14.35 ATOM 2967 O GLY A 387 −21.791 26.644−14.129 1.00 13.79 ATOM 2968 N PHE A 388 −21.771 24.453 −13.624 1.0014.30 ATOM 2969 CA PHE A 388 −20.479 24.217 −14.312 1.00 14.69 ATOM 2970CB PHE A 388 −19.912 22.824 −13.987 1.00 14.42 ATOM 2971 CG PHE A 388−19.359 22.730 −12.584 1.00 13.79 ATOM 2972 CD1 PHE A 388 −18.139 23.335−12.269 1.00 14.50 ATOM 2973 CE1 PHE A 388 −17.621 23.261 −10.947 1.0016.38 ATOM 2974 CZ PHE A 388 −18.377 22.627 −9.951 1.00 14.77 ATOM 2975CE2 PHE A 388 −19.604 22.045 −10.265 1.00 16.35 ATOM 2976 CD2 PHE A 388−20.088 22.100 −11.578 1.00 14.00 ATOM 2977 C PHE A 388 −20.601 24.428−15.821 1.00 15.31 ATOM 2978 O PHE A 388 −19.740 25.078 −16.440 1.0015.55 ATOM 2979 N LEU A 389 −21.669 23.913 −16.415 1.00 16.52 ATOM 2980CA LEU A 389 −21.889 24.159 −17.856 1.00 17.84 ATOM 2981 CB LEU A 389−23.137 23.431 −18.382 1.00 17.83 ATOM 2982 CG LEU A 389 −23.172 21.911−18.427 1.00 22.75 ATOM 2983 CD1 LEU A 389 −24.247 21.418 −19.401 1.0022.55 ATOM 2984 CD2 LEU A 389 −21.805 21.333 −18.806 1.00 24.63 ATOM2985 C LEU A 389 −22.013 25.634 −18.136 1.00 18.32 ATOM 2986 O LEU A 389−21.409 26.138 −19.091 1.00 19.50 ATOM 2987 N SER A 390 −22.775 26.341−17.295 1.00 18.55 ATOM 2988 CA SER A 390 −23.021 27.767 −17.469 1.0019.33 ATOM 2989 CB SER A 390 −24.090 28.246 −16.491 1.00 19.89 ATOM 2990OG SER A 390 −25.325 27.693 −16.891 1.00 24.07 ATOM 2991 C SER A 390−21.763 28.603 −17.323 1.00 19.51 ATOM 2992 O SER A 390 −21.575 29.585−18.055 1.00 19.69 ATOM 2993 N GLU A 391 −20.893 28.200 −16.399 1.0018.87 ATOM 2994 CA GLU A 391 −19.633 28.879 −16.220 1.00 19.59 ATOM 2995CB GLU A 391 −18.901 28.393 −14.952 1.00 20.02 ATOM 2996 CG GLU A 391−19.528 28.924 −13.668 1.00 23.54 ATOM 2997 CD GLU A 391 −19.590 30.448−13.634 1.00 26.61 ATOM 2998 OE1 GLU A 391 −18.609 31.102 −14.023 1.0028.42 ATOM 2999 OE2 GLU A 391 −20.637 30.994 −13.227 1.00 29.52 ATOM3000 C GLU A 391 −18.738 28.729 −17.457 1.00 19.12 ATOM 3001 O GLU A 391−18.123 29.709 −17.906 1.00 19.40 ATOM 3002 N ALA A 392 −18.654 27.516−17.991 1.00 18.81 ATOM 3003 CA ALA A 392 −17.861 27.304 −19.201 1.0019.72 ATOM 3004 CB ALA A 392 −17.758 25.815 −19.526 1.00 19.45 ATOM 3005C ALA A 392 −18.478 28.098 −20.363 1.00 19.59 ATOM 3006 O ALA A 392−17.764 28.724 −21.157 1.00 18.65 ATOM 3007 N ALA A 393 −19.808 28.117−20.425 1.00 20.40 ATOM 3008 CA ALA A 393 −20.526 28.820 −21.507 1.0021.27 ATOM 3009 CB ALA A 393 −22.035 28.524 −21.421 1.00 21.83 ATOM 3010C ALA A 393 −20.266 30.334 −21.574 1.00 21.79 ATOM 3011 O ALA A 393−20.283 30.920 −22.664 1.00 21.96 ATOM 3012 N LYS A 394 −19.976 30.971−20.435 1.00 21.37 ATOM 3013 CA LYS A 394 −19.626 32.383 −20.447 1.0022.02 ATOM 3014 CB LYS A 394 −19.289 32.916 −19.043 1.00 23.11 ATOM 3015CG LYS A 394 −20.411 32.980 −18.044 1.00 25.98 ATOM 3016 CD LYS A 394−19.782 33.341 −16.700 1.00 28.89 ATOM 3017 CE LYS A 394 −20.793 33.314−15.576 1.00 34.05 ATOM 3018 NZ LYS A 394 −20.097 33.674 −14.290 1.0033.96 ATOM 3019 C LYS A 394 −18.403 32.626 −21.310 1.00 21.55 ATOM 3020O LYS A 394 −18.188 33.742 −21.771 1.00 22.41 ATOM 3021 N TYR A 395−17.570 31.604 −21.488 1.00 19.98 ATOM 3022 CA TYR A 395 −16.287 31.824−22.132 1.00 19.46 ATOM 3023 CB TYR A 395 −15.137 31.464 −21.185 1.0020.82 ATOM 3024 CG TYR A 395 −15.291 32.165 −19.872 1.00 22.04 ATOM 3025CD1 TYR A 395 −15.644 31.450 −18.716 1.00 23.21 ATOM 3026 CE1 TYR A 395−15.806 32.097 −17.508 1.00 23.70 ATOM 3027 CZ TYR A 395 −15.661 33.473−17.460 1.00 23.93 ATOM 3028 OH TYR A 395 −15.828 34.143 −16.272 1.0026.14 ATOM 3029 CE2 TYR A 395 −15.327 34.202 −18.593 1.00 24.38 ATOM3030 CD2 TYR A 395 −15.157 33.548 −19.791 1.00 22.37 ATOM 3031 C TYR A395 −16.157 31.119 −23.451 1.00 19.22 ATOM 3032 O TYR A 395 −15.04530.940 −23.941 1.00 18.70 ATOM 3033 N VAL A 396 −17.299 30.718 −24.0181.00 18.16 ATOM 3034 CA VAL A 396 −17.331 30.135 −25.352 1.00 19.03 ATOM3035 CB VAL A 396 −18.396 29.025 −25.458 1.00 18.18 ATOM 3036 CG1 VAL A396 −18.469 28.465 −26.898 1.00 18.63 ATOM 3037 CG2 VAL A 396 −18.09427.915 −24.452 1.00 18.97 ATOM 3038 C VAL A 396 −17.654 31.288 −26.3081.00 19.65 ATOM 3039 O VAL A 396 −18.644 31.986 −26.098 1.00 19.66 ATOM3040 N PRO A 397 −16.810 31.507 −27.328 1.00 20.41 ATOM 3041 CA PRO A397 −17.016 32.626 −28.256 1.00 20.82 ATOM 3042 CB PRO A 397 −15.79432.561 −29.175 1.00 21.47 ATOM 3043 CG PRO A 397 −14.819 31.725 −28.4751.00 21.69 ATOM 3044 CD PRO A 397 −15.598 30.741 −27.661 1.00 19.70 ATOM3045 C PRO A 397 −18.280 32.434 −29.073 1.00 21.11 ATOM 3046 O PRO A 397−18.844 31.339 −29.088 1.00 19.88 ATOM 3047 N ALA A 398 −18.713 33.492−29.765 1.00 21.26 ATOM 3048 CA ALA A 398 −19.951 33.424 −30.559 1.0021.44 ATOM 3049 CB ALA A 398 −20.227 34.766 −31.230 1.00 22.38 ATOM 3050C ALA A 398 −19.971 32.297 −31.587 1.00 21.30 ATOM 3051 O ALA A 398−21.038 31.769 −31.901 1.00 22.15 ATOM 3052 N ASP A 399 −18.804 31.896−32.102 1.00 20.30 ATOM 3053 CA ASP A 399 −18.780 30.858 −33.133 1.0019.40 ATOM 3054 CB ASP A 399 −17.587 31.032 −34.071 1.00 19.42 ATOM 3055CG ASP A 399 −16.233 30.835 −33.381 1.00 21.84 ATOM 3056 OD1 ASP A 399−16.146 30.569 −32.159 1.00 20.91 ATOM 3057 OD2 ASP A 399 −15.229 30.950−34.104 1.00 24.62 ATOM 3058 C ASP A 399 −18.834 29.435 −32.579 1.0018.14 ATOM 3059 O ASP A 399 −18.802 28.465 −33.350 1.00 16.78 ATOM 3060N GLY A 400 −18.891 29.322 −31.245 1.00 16.82 ATOM 3061 CA GLY A 400−18.996 28.015 −30.607 1.00 15.41 ATOM 3062 C GLY A 400 −17.693 27.229−30.556 1.00 15.07 ATOM 3063 O GLY A 400 −17.704 26.041 −30.203 1.0015.23 ATOM 3064 N SER A 401 −16.572 27.861 −30.882 1.00 14.21 ATOM 3065CA SER A 401 −15.312 27.119 −30.893 1.00 14.76 ATOM 3066 CB SER A 401−14.241 27.840 −31.718 1.00 14.71 ATOM 3067 OG SER A 401 −14.059 29.160−31.257 1.00 16.86 ATOM 3068 C SER A 401 −14.815 26.866 −29.448 1.0014.38 ATOM 3069 O SER A 401 −14.992 27.717 −28.562 1.00 14.58 ATOM 3070N LEU A 402 −14.169 25.720 −29.249 1.00 13.54 ATOM 3071 CA LEU A 402−13.603 25.364 −27.968 1.00 13.21 ATOM 3072 CB LEU A 402 −14.271 24.080−27.450 1.00 13.42 ATOM 3073 CG LEU A 402 −15.776 24.192 −27.162 1.0014.06 ATOM 3074 CD1 LEU A 402 −16.289 22.834 −26.668 1.00 13.87 ATOM3075 CD2 LEU A 402 −15.997 25.264 −26.109 1.00 17.00 ATOM 3076 C LEU A402 −12.111 25.143 −28.109 1.00 12.99 ATOM 3077 O LEU A 402 −11.69524.166 −28.707 1.00 13.19 ATOM 3078 N ALA A 403 −11.320 26.070 −27.5781.00 13.02 ATOM 3079 CA ALA A 403 −9.884 25.870 −27.454 1.00 12.06 ATOM3080 CB ALA A 403 −9.194 27.226 −27.220 1.00 11.77 ATOM 3081 C ALA A 403−9.591 24.907 −26.300 1.00 12.81 ATOM 3082 O ALA A 403 −10.508 24.315−25.714 1.00 12.36 ATOM 3083 N GLU A 404 −8.308 24.772 −25.959 1.0011.39 ATOM 3084 CA GLU A 404 −7.918 23.922 −24.855 1.00 11.47 ATOM 3085CB GLU A 404 −6.412 23.689 −24.931 1.00 11.02 ATOM 3086 CG GLU A 404−5.865 22.723 −23.873 1.00 11.30 ATOM 3087 CD GLU A 404 −4.363 22.669−23.954 1.00 12.25 ATOM 3088 OE1 GLU A 404 −3.729 23.692 −23.622 1.0012.06 ATOM 3089 OE2 GLU A 404 −3.818 21.624 −24.390 1.00 12.60 ATOM 3090C GLU A 404 −8.246 24.635 −23.538 1.00 11.77 ATOM 3091 O GLU A 404−8.755 24.006 −22.590 1.00 11.08 ATOM 3092 N GLN A 405 −7.890 25.924−23.453 1.00 12.07 ATOM 3093 CA GLN A 405 −7.952 26.655 −22.196 1.0013.33 ATOM 3094 CB GLN A 405 −6.539 26.986 −21.678 1.00 13.24 ATOM 3095CG GLN A 405 −5.625 25.821 −21.553 1.00 15.96 ATOM 3096 CD GLN A 405−4.229 26.213 −21.051 1.00 14.97 ATOM 3097 OE1 GLN A 405 −4.068 27.130−20.236 1.00 15.09 ATOM 3098 NE2 GLN A 405 −3.236 25.475 −21.496 1.0015.87 ATOM 3099 C GLN A 405 −8.687 27.985 −22.356 1.00 13.51 ATOM 3100 OGLN A 405 −8.870 28.475 −23.489 1.00 13.74 ATOM 3101 N PHE A 406 −9.04128.587 −21.226 1.00 12.60 ATOM 3102 CA PHE A 406 −9.516 29.990 −21.2071.00 13.82 ATOM 3103 CB PHE A 406 −11.058 30.104 −21.232 1.00 13.56 ATOM3104 CG PHE A 406 −11.800 29.385 −20.123 1.00 15.20 ATOM 3105 CD1 PHE A406 −12.155 28.026 −20.242 1.00 16.33 ATOM 3106 CE1 PHE A 406 −12.87927.370 −19.239 1.00 16.80 ATOM 3107 CZ PHE A 406 −13.340 28.094 −18.1141.00 16.74 ATOM 3108 CE2 PHE A 406 −13.020 29.453 −17.993 1.00 15.37ATOM 3109 CD2 PHE A 406 −12.260 30.101 −19.000 1.00 17.56 ATOM 3110 CPHE A 406 −8.836 30.737 −20.078 1.00 13.80 ATOM 3111 O PHE A 406 −8.58730.151 −19.018 1.00 14.02 ATOM 3112 N ASP A 407 −8.481 31.997 −20.3211.00 14.98 ATOM 3113 CA ASP A 407 −7.547 32.726 −19.438 1.00 15.04 ATOM3114 CB ASP A 407 −7.237 34.115 −20.032 1.00 15.99 ATOM 3115 CG ASP A407 −6.159 34.829 −19.293 1.00 18.17 ATOM 3116 OD1 ASP A 407 −6.47435.508 −18.293 1.00 20.41 ATOM 3117 OD2 ASP A 407 −4.993 34.683 −19.6851.00 20.75 ATOM 3118 C ASP A 407 −8.100 32.829 −18.005 1.00 14.54 ATOM3119 O ASP A 407 −9.257 33.185 −17.800 1.00 15.12 ATOM 3120 N ARG A 408−7.248 32.548 −17.018 1.00 14.63 ATOM 3121 CA ARG A 408 −7.644 32.530−15.609 1.00 15.33 ATOM 3122 CB ARG A 408 −6.453 32.095 −14.753 1.0015.32 ATOM 3123 CG ARG A 408 −5.236 33.062 −14.828 1.00 14.48 ATOM 3124CD ARG A 408 −4.009 32.479 −14.122 1.00 16.22 ATOM 3125 NE ARG A 408−4.237 32.248 −12.695 1.00 15.55 ATOM 3126 CZ ARG A 408 −3.613 31.323−11.961 1.00 18.66 ATOM 3127 NH1 ARG A 408 −3.878 31.231 −10.658 1.0017.51 ATOM 3128 NH2 ARG A 408 −2.717 30.499 −12.511 1.00 17.12 ATOM 3129C ARG A 408 −8.167 33.886 −15.108 1.00 16.66 ATOM 3130 O ARG A 408−8.898 33.943 −14.110 1.00 16.82 ATOM 3131 N ASN A 409 −7.781 34.964−15.790 1.00 18.00 ATOM 3132 CA ASN A 409 −8.252 36.316 −15.421 1.0020.06 ATOM 3133 CB ASN A 409 −7.069 37.275 −15.355 1.00 20.32 ATOM 3134CG ASN A 409 −6.119 36.937 −14.224 1.00 21.65 ATOM 3135 OD1 ASN A 409−6.549 36.678 −13.111 1.00 24.42 ATOM 3136 ND2 ASN A 409 −4.830 36.914−14.516 1.00 23.96 ATOM 3137 C ASN A 409 −9.320 36.903 −16.336 1.0021.43 ATOM 3138 O ASN A 409 −10.272 37.524 −15.857 1.00 22.03 ATOM 3139N SER A 410 −9.152 36.724 −17.646 1.00 22.25 ATOM 3140 CA SER A 410−10.007 37.410 −18.624 1.00 22.76 ATOM 3141 CB SER A 410 −9.146 38.159−19.636 1.00 23.45 ATOM 3142 OG SER A 410 −8.470 37.260 −20.495 1.0023.38 ATOM 3143 C SER A 410 −10.971 36.472 −19.343 1.00 22.82 ATOM 3144O SER A 410 −11.898 36.925 −20.010 1.00 23.59 ATOM 3145 N GLY A 411−10.758 35.161 −19.238 1.00 21.64 ATOM 3146 CA GLY A 411 −11.668 34.221−19.877 1.00 20.39 ATOM 3147 C GLY A 411 −11.476 34.087 −21.379 1.0019.99 ATOM 3148 O GLY A 411 −12.223 33.368 −22.029 1.00 20.85 ATOM 3149N THR A 412 −10.478 34.750 −21.941 1.00 19.04 ATOM 3150 CA THR A 412−10.268 34.658 −23.383 1.00 20.25 ATOM 3151 CB THR A 412 −9.447 35.853−23.922 1.00 21.81 ATOM 3152 OG1 THR A 412 −8.187 35.900 −23.257 1.0026.24 ATOM 3153 CG2 THR A 412 −10.160 37.163 −23.631 1.00 23.82 ATOM3154 C THR A 412 −9.615 33.294 −23.732 1.00 19.17 ATOM 3155 O THR A 412−8.786 32.796 −22.970 1.00 17.70 ATOM 3156 N PRO A 413 −9.996 32.688−24.874 1.00 18.84 ATOM 3157 CA PRO A 413 −9.466 31.348 −25.234 1.0018.32 ATOM 3158 CB PRO A 413 −10.220 31.002 −26.525 1.00 18.75 ATOM 3159CG PRO A 413 −11.513 31.928 −26.451 1.00 19.42 ATOM 3160 CD PRO A 413−10.943 33.195 −25.891 1.00 19.40 ATOM 3161 C PRO A 413 −7.959 31.399−25.464 1.00 18.87 ATOM 3162 O PRO A 413 −7.436 32.406 −25.955 1.0018.20 ATOM 3163 N LEU A 414 −7.253 30.353 −25.051 1.00 18.44 ATOM 3164CA LEU A 414 −5.822 30.275 −25.305 1.00 18.90 ATOM 3165 CB LEU A 414−4.992 30.974 −24.208 1.00 21.66 ATOM 3166 CG LEU A 414 −5.019 30.574−22.754 1.00 24.35 ATOM 3167 CD1 LEU A 414 −4.134 31.484 −21.892 1.0027.42 ATOM 3168 CD2 LEU A 414 −6.406 30.669 −22.224 1.00 30.77 ATOM 3169C LEU A 414 −5.362 28.854 −25.518 1.00 17.24 ATOM 3170 O LEU A 414−6.138 27.913 −25.406 1.00 15.97 ATOM 3171 N SER A 415 −4.091 28.733−25.865 1.00 15.34 ATOM 3172 CA SER A 415 −3.473 27.481 −26.257 1.0015.28 ATOM 3173 CB SER A 415 −3.434 26.468 −25.101 1.00 14.94 ATOM 3174OG SER A 415 −2.632 25.355 −25.445 1.00 14.00 ATOM 3175 C SER A 415−4.141 26.932 −27.528 1.00 15.13 ATOM 3176 O SER A 415 −4.665 27.718−28.334 1.00 14.92 ATOM 3177 N ALA A 416 −4.097 25.618 −27.714 1.0014.27 ATOM 3178 CA ALA A 416 −4.540 24.977 −28.976 1.00 14.25 ATOM 3179CB ALA A 416 −4.380 23.486 −28.889 1.00 13.68 ATOM 3180 C ALA A 416−5.981 25.314 −29.315 1.00 14.33 ATOM 3181 O ALA A 416 −6.854 25.216−28.459 1.00 14.09 ATOM 3182 N LEU A 417 −6.223 25.680 −30.567 1.0013.34 ATOM 3183 CA LEU A 417 −7.584 25.985 −31.006 1.00 14.22 ATOM 3184CB LEU A 417 −7.536 26.931 −32.194 1.00 16.32 ATOM 3185 CG LEU A 417−6.841 28.283 −31.942 1.00 18.75 ATOM 3186 CD1 LEU A 417 −7.005 29.127−33.163 1.00 23.81 ATOM 3187 CD2 LEU A 417 −7.419 28.991 −30.712 1.0021.76 ATOM 3188 C LEU A 417 −8.279 24.687 −31.413 1.00 13.18 ATOM 3189 OLEU A 417 −7.610 23.712 −31.775 1.00 13.19 ATOM 3190 N HIS A 418 −9.60924.658 −31.311 1.00 12.26 ATOM 3191 CA HIS A 418 −10.399 23.496 −31.7641.00 11.77 ATOM 3192 CB HIS A 418 −10.487 23.454 −33.303 1.00 13.41 ATOM3193 CG HIS A 418 −11.294 24.566 −33.898 1.00 14.68 ATOM 3194 ND1 HIS A418 −12.646 24.717 −33.660 1.00 16.00 ATOM 3195 CE1 HIS A 418 −13.09525.762 −34.341 1.00 17.83 ATOM 3196 NE2 HIS A 418 −12.085 26.290 −35.0151.00 17.38 ATOM 3197 CD2 HIS A 418 −10.948 25.557 −34.763 1.00 17.65ATOM 3198 C HIS A 418 −9.826 22.187 −31.206 1.00 12.24 ATOM 3199 O HIS A418 −9.540 21.250 −31.947 1.00 12.10 ATOM 3200 N LEU A 419 −9.656 22.116−29.880 1.00 10.99 ATOM 3201 CA LEU A 419 −9.152 20.881 −29.301 1.0010.46 ATOM 3202 CB LEU A 419 −8.742 21.069 −27.826 1.00 10.91 ATOM 3203CG LEU A 419 −7.983 19.883 −27.220 1.00 10.16 ATOM 3204 CD1 LEU A 419−6.524 19.944 −27.669 1.00 11.45 ATOM 3205 CD2 LEU A 419 −8.080 19.960−25.657 1.00 10.73 ATOM 3206 C LEU A 419 −10.215 19.812 −29.398 1.0010.49 ATOM 3207 O LEU A 419 −11.312 19.973 −28.863 1.00 10.67 ATOM 3208N THR A 420 −9.860 18.686 −30.021 1.00 10.22 ATOM 3209 CA THR A 420−10.833 17.629 −30.296 1.00 10.90 ATOM 3210 CB THR A 420 −10.201 16.460−31.096 1.00 11.55 ATOM 3211 OG1 THR A 420 −9.357 16.999 −32.115 1.0012.00 ATOM 3212 CG2 THR A 420 −11.310 15.625 −31.786 1.00 12.52 ATOM3213 C THR A 420 −11.426 17.135 −28.995 1.00 11.43 ATOM 3214 O THR A 420−12.648 16.903 −28.891 1.00 11.83 ATOM 3215 N TRP A 421 −10.569 16.987−27.980 1.00 9.79 ATOM 3216 CA TRP A 421 −11.052 16.511 −26.688 1.0011.67 ATOM 3217 CB TRP A 421 −9.839 16.184 −25.803 1.00 12.06 ATOM 3218CG TRP A 421 −10.075 15.476 −24.508 1.00 11.94 ATOM 3219 CD1 TRP A 421−11.274 15.202 −23.881 1.00 14.46 ATOM 3220 NE1 TRP A 421 −11.044 14.590−22.663 1.00 14.38 ATOM 3221 CE2 TRP A 421 −9.691 14.497 −22.468 1.0013.52 ATOM 3222 CD2 TRP A 421 −9.053 15.039 −23.616 1.00 13.01 ATOM 3223CE3 TRP A 421 −7.652 15.045 −23.680 1.00 15.38 ATOM 3224 CZ3 TRP A 421−6.932 14.503 −22.605 1.00 16.10 ATOM 3225 CH2 TRP A 421 −7.603 13.973−21.475 1.00 14.87 ATOM 3226 CZ2 TRP A 421 −8.973 13.945 −21.398 1.0014.27 ATOM 3227 C TRP A 421 −12.035 17.514 −26.032 1.00 10.89 ATOM 3228O TRP A 421 −12.966 17.092 −25.357 1.00 10.96 ATOM 3229 N SER A 422−11.844 18.822 −26.211 1.00 10.49 ATOM 3230 CA SER A 422 −12.833 19.794−25.696 1.00 11.13 ATOM 3231 CB SER A 422 −12.459 21.243 −26.049 1.0010.97 ATOM 3232 OG SER A 422 −11.302 21.682 −25.338 1.00 13.73 ATOM 3233C SER A 422 −14.229 19.496 −26.257 1.00 11.40 ATOM 3234 O SER A 422−15.204 19.468 −25.530 1.00 12.11 ATOM 3235 N TYR A 423 −14.320 19.281−27.563 1.00 11.29 ATOM 3236 CA TYR A 423 −15.617 18.990 −28.170 1.0011.29 ATOM 3237 CB TYR A 423 −15.502 19.020 −29.717 1.00 11.85 ATOM 3238CG TYR A 423 −15.132 20.389 −30.274 1.00 12.18 ATOM 3239 CD1 TYR A 423−16.002 21.485 −30.145 1.00 10.35 ATOM 3240 CE1 TYR A 423 −15.668 22.741−30.643 1.00 11.83 ATOM 3241 CZ TYR A 423 −14.468 22.912 −31.316 1.0012.29 ATOM 3242 OH TYR A 423 −14.157 24.145 −31.783 1.00 13.08 ATOM 3243CE2 TYR A 423 −13.588 21.845 −31.496 1.00 13.57 ATOM 3244 CD2 TYR A 423−13.942 20.572 −30.991 1.00 12.74 ATOM 3245 C TYR A 423 −16.217 17.673−27.658 1.00 12.12 ATOM 3246 O TYR A 423 −17.430 17.622 −27.323 1.0012.13 ATOM 3247 N ALA A 424 −15.385 16.623 −27.539 1.00 10.99 ATOM 3248CA ALA A 424 −15.853 15.337 −26.986 1.00 11.26 ATOM 3249 CB ALA A 424−14.717 14.294 −26.952 1.00 11.73 ATOM 3250 C ALA A 424 −16.411 15.535−25.588 1.00 10.93 ATOM 3251 O ALA A 424 −17.465 14.974 −25.246 1.0011.68 ATOM 3252 N SER A 425 −15.696 16.308 −24.770 1.00 9.74 ATOM 3253CA SER A 425 −16.077 16.519 −23.379 1.00 10.92 ATOM 3254 CB SER A 425−14.948 17.228 −22.602 1.00 11.59 ATOM 3255 OG SER A 425 −14.817 18.604−22.957 1.00 13.86 ATOM 3256 C SER A 425 −17.402 17.291 −23.219 1.0011.65 ATOM 3257 O SER A 425 −18.132 17.043 −22.276 1.00 11.86 ATOM 3258N PHE A 426 −17.683 18.220 −24.133 1.00 12.13 ATOM 3259 CA PHE A 426−18.983 18.891 −24.154 1.00 12.95 ATOM 3260 CB PHE A 426 −19.018 20.053−25.173 1.00 13.14 ATOM 3261 CG PHE A 426 −20.410 20.575 −25.391 1.0015.93 ATOM 3262 CD1 PHE A 426 −20.951 21.522 −24.516 1.00 17.76 ATOM3263 CE1 PHE A 426 −22.279 21.980 −24.688 1.00 17.49 ATOM 3264 CZ PHE A426 −23.064 21.455 −25.716 1.00 17.22 ATOM 3265 CE2 PHE A 426 −22.56120.496 −26.558 1.00 17.04 ATOM 3266 CD2 PHE A 426 −21.225 20.045 −26.3961.00 17.29 ATOM 3267 C PHE A 426 −20.079 17.878 −24.502 1.00 13.31 ATOM3268 O PHE A 426 −21.120 17.827 −23.850 1.00 13.38 ATOM 3269 N LEU A 427−19.834 17.077 −25.539 1.00 13.23 ATOM 3270 CA LEU A 427 −20.811 16.090−25.996 1.00 14.41 ATOM 3271 CB LEU A 427 −20.339 15.410 −27.291 1.0014.32 ATOM 3272 CG LEU A 427 −20.363 16.336 −28.506 1.00 15.96 ATOM 3273CD1 LEU A 427 −19.661 15.639 −29.689 1.00 18.66 ATOM 3274 CD2 LEU A 427−21.773 16.800 −28.876 1.00 16.20 ATOM 3275 C LEU A 427 −21.137 15.045−24.959 1.00 14.86 ATOM 3276 O LEU A 427 −22.307 14.667 −24.833 1.0016.01 ATOM 3277 N THR A 428 −20.130 14.551 −24.235 1.00 13.81 ATOM 3278CA THR A 428 −20.397 13.544 −23.196 1.00 13.67 ATOM 3279 CB THR A 428−19.134 12.732 −22.745 1.00 12.87 ATOM 3280 OG1 THR A 428 −18.127 13.597−22.185 1.00 12.33 ATOM 3281 CG2 THR A 428 −18.533 11.902 −23.923 1.0013.77 ATOM 3282 C THR A 428 −21.146 14.133 −21.980 1.00 13.66 ATOM 3283O THR A 428 −22.102 13.517 −21.478 1.00 15.09 ATOM 3284 N ALA A 429−20.738 15.312 −21.524 1.00 13.72 ATOM 3285 CA ALA A 429 −21.367 15.942−20.355 1.00 14.65 ATOM 3286 CB ALA A 429 −20.704 17.295 −20.036 1.0014.11 ATOM 3287 C ALA A 429 −22.852 16.158 −20.643 1.00 15.47 ATOM 3288O ALA A 429 −23.704 15.908 −19.783 1.00 15.97 ATOM 3289 N THR A 430−23.151 16.622 −21.854 1.00 15.48 ATOM 3290 CA THR A 430 −24.554 16.940−22.208 1.00 15.64 ATOM 3291 CB THR A 430 −24.675 17.968 −23.353 1.0016.50 ATOM 3292 OG1 THR A 430 −23.980 17.494 −24.514 1.00 16.24 ATOM3293 CG2 THR A 430 −24.101 19.317 −22.916 1.00 16.43 ATOM 3294 C THR A430 −25.401 15.690 −22.463 1.00 15.41 ATOM 3295 O THR A 430 −26.61115.674 −22.158 1.00 15.67 ATOM 3296 N ALA A 431 −24.772 14.632 −22.9681.00 15.22 ATOM 3297 CA ALA A 431 −25.437 13.311 −23.049 1.00 16.26 ATOM3298 CB ALA A 431 −24.560 12.300 −23.762 1.00 16.39 ATOM 3299 C ALA A431 −25.842 12.797 −21.673 1.00 16.53 ATOM 3300 O ALA A 431 −26.98512.319 −21.485 1.00 16.61 ATOM 3301 N ARG A 432 −24.937 12.895 −20.6891.00 15.66 ATOM 3302 CA ARG A 432 −25.256 12.410 −19.342 1.00 15.64 ATOM3303 CB ARG A 432 −24.022 12.413 −18.432 1.00 15.83 ATOM 3304 CG ARG A432 −22.862 11.574 −18.994 1.00 14.85 ATOM 3305 CD ARG A 432 −23.17410.051 −19.154 1.00 17.10 ATOM 3306 NE ARG A 432 −21.958 9.472 −19.7081.00 18.67 ATOM 3307 CZ ARG A 432 −21.766 9.225 −21.003 1.00 20.81 ATOM3308 NH1 ARG A 432 −22.769 9.372 −21.868 1.00 17.45 ATOM 3309 NH2 ARG A432 −20.576 8.781 −21.427 1.00 19.90 ATOM 3310 C ARG A 432 −26.37513.235 −18.719 1.00 16.42 ATOM 3311 O ARG A 432 −27.256 12.685 −18.0301.00 17.47 ATOM 3312 N ARG A 433 −26.371 14.535 −18.996 1.00 16.54 ATOM3313 CA ARG A 433 −27.425 15.418 −18.493 1.00 17.60 ATOM 3314 CB ARG A433 −27.204 16.852 −18.960 1.00 16.74 ATOM 3315 CG ARG A 433 −28.28717.833 −18.461 1.00 18.39 ATOM 3316 CD ARG A 433 −27.931 19.239 −18.8661.00 20.79 ATOM 3317 NE ARG A 433 −28.739 20.260 −18.166 1.00 23.05 ATOM3318 CZ ARG A 433 −29.859 20.799 −18.654 1.00 26.97 ATOM 3319 NH1 ARG A433 −30.333 20.404 −19.837 1.00 24.94 ATOM 3320 NH2 ARG A 433 −30.50621.738 −17.954 1.00 25.73 ATOM 3321 C ARG A 433 −28.793 14.943 −18.9561.00 17.90 ATOM 3322 O ARG A 433 −29.761 14.984 −18.184 1.00 18.50 ATOM3323 N ALA A 434 −28.861 14.502 −20.210 1.00 18.06 ATOM 3324 CA ALA A434 −30.103 13.993 −20.806 1.00 19.10 ATOM 3325 CB ALA A 434 −30.06714.202 −22.318 1.00 19.95 ATOM 3326 C ALA A 434 −30.399 12.532 −20.4751.00 20.48 ATOM 3327 O ALA A 434 −31.371 11.975 −20.980 1.00 21.63 ATOM3328 N GLY A 435 −29.594 11.904 −19.620 1.00 19.94 ATOM 3329 CA GLY A435 −29.865 10.531 −19.182 1.00 21.09 ATOM 3330 C GLY A 435 −29.4159.475 −20.188 1.00 21.57 ATOM 3331 O GLY A 435 −29.847 8.304 −20.1301.00 21.64 ATOM 3332 N ILE A 436 −28.529 9.873 −21.100 1.00 20.44 ATOM3333 CA ILE A 436 −27.951 8.937 −22.060 1.00 20.81 ATOM 3334 CB ILE A436 −27.753 9.601 −23.447 1.00 20.75 ATOM 3335 CG1 ILE A 436 −29.1329.977 −24.027 1.00 23.03 ATOM 3336 CD1 ILE A 436 −29.103 11.031 −25.1281.00 26.34 ATOM 3337 CG2 ILE A 436 −27.031 8.643 −24.395 1.00 21.35 ATOM3338 C ILE A 436 −26.634 8.412 −21.485 1.00 20.99 ATOM 3339 O ILE A 436−25.666 9.171 −21.339 1.00 20.46 ATOM 3340 N VAL A 437 −26.616 7.120−21.162 1.00 20.70 ATOM 3341 CA VAL A 437 −25.465 6.479 −20.517 1.0021.39 ATOM 3342 CB VAL A 437 −25.848 5.826 −19.160 1.00 21.74 ATOM 3343CG1 VAL A 437 −26.340 6.911 −18.205 1.00 22.12 ATOM 3344 CG2 VAL A 437−26.909 4.703 −19.334 1.00 21.83 ATOM 3345 C VAL A 437 −24.802 5.459−21.444 1.00 21.79 ATOM 3346 O VAL A 437 −25.459 4.901 −22.312 1.0022.18 ATOM 3347 N PRO A 438 −23.497 5.208 −21.255 1.00 22.28 ATOM 3348CA PRO A 438 −22.837 4.291 −22.181 1.00 22.83 ATOM 3349 CB PRO A 438−21.365 4.642 −22.009 1.00 22.45 ATOM 3350 CG PRO A 438 −21.248 5.054−20.578 1.00 23.88 ATOM 3351 CD PRO A 438 −22.575 5.707 −20.214 1.0022.50 ATOM 3352 C PRO A 438 −23.093 2.840 −21.753 1.00 22.80 ATOM 3353 OPRO A 438 −23.580 2.604 −20.626 1.00 23.06 ATOM 3354 N PRO A 439 −22.7961.878 −22.639 1.00 22.65 ATOM 3355 CA PRO A 439 −22.911 0.452 −22.2831.00 22.08 ATOM 3356 CB PRO A 439 −22.300 −0.269 −23.499 1.00 21.30 ATOM3357 CG PRO A 439 −22.526 0.664 −24.618 1.00 22.81 ATOM 3358 CD PRO A439 −22.391 2.062 −24.050 1.00 22.29 ATOM 3359 C PRO A 439 −22.122 0.129−21.037 1.00 21.88 ATOM 3360 O PRO A 439 −21.075 0.750 −20.776 1.0020.95 ATOM 3361 N SER A 440 −22.628 −0.818 −20.253 1.00 22.34 ATOM 3362CA SER A 440 −21.932 −1.273 −19.060 1.00 23.42 ATOM 3363 CB SER A 440−22.818 −2.195 −18.224 1.00 24.01 ATOM 3364 OG SER A 440 −23.805 −1.412−17.566 1.00 26.78 ATOM 3365 C SER A 440 −20.654 −1.992 −19.430 1.0023.78 ATOM 3366 O SER A 440 −20.540 −2.554 −20.522 1.00 23.97 ATOM 3367N TRP A 441 −19.681 −1.929 −18.536 1.00 24.15 ATOM 3368 CA TRP A 441−18.431 −2.646 −18.718 1.00 24.63 ATOM 3369 CB TRP A 441 −17.255 −1.679−18.819 1.00 22.62 ATOM 3370 CG TRP A 441 −16.963 −0.837 −17.583 1.0019.24 ATOM 3371 CD1 TRP A 441 −17.409 0.432 −17.339 1.00 16.95 ATOM 3372NE1 TRP A 441 −16.909 0.878 −16.138 1.00 17.89 ATOM 3373 CE2 TRP A 441−16.111 −0.098 −15.595 1.00 17.88 ATOM 3374 CD2 TRP A 441 −16.130 −1.194−16.476 1.00 18.22 ATOM 3375 CE3 TRP A 441 −15.377 −2.338 −16.149 1.0019.64 ATOM 3376 CZ3 TRP A 441 −14.658 −2.355 −14.961 1.00 18.61 ATOM3377 CH2 TRP A 441 −14.670 −1.246 −14.094 1.00 20.62 ATOM 3378 CZ2 TRP A441 −15.400 −0.114 −14.391 1.00 19.39 ATOM 3379 C TRP A 441 −18.179−3.661 −17.625 1.00 26.71 ATOM 3380 O TRP A 441 −17.410 −4.592 −17.8131.00 25.60 ATOM 3381 N ALA A 442 −18.798 −3.471 −16.468 1.00 29.78 ATOM3382 CA ALA A 442 −18.442 −4.292 −15.330 1.00 33.94 ATOM 3383 CB ALA A442 −18.347 −3.447 −14.082 1.00 33.18 ATOM 3384 C ALA A 442 −19.447−5.412 −15.136 1.00 37.28 ATOM 3385 O ALA A 442 −20.383 −5.561 −15.9151.00 38.51 ATOM 3386 N ASN A 443 −19.201 −6.222 −14.116 1.00 41.45 ATOM3387 CA ASN A 443 −20.226 −7.030 −13.467 1.00 45.10 ATOM 3388 CB ASN A443 −20.135 −8.490 −13.914 1.00 45.75 ATOM 3389 CG ASN A 443 −18.815−9.129 −13.531 1.00 48.14 ATOM 3390 OD1 ASN A 443 −18.620 −9.524 −12.3801.00 50.51 ATOM 3391 ND2 ASN A 443 −17.888 −9.212 −14.492 1.00 50.01ATOM 3392 C ASN A 443 −19.946 −6.892 −11.972 1.00 46.93 ATOM 3393 O ASNA 443 −18.878 −6.396 −11.580 1.00 47.13 ATOM 3394 N SER A 444 −20.873−7.343 −11.135 1.00 49.21 ATOM 3395 CA SER A 444 −20.719 −7.201 −9.6841.00 51.08 ATOM 3396 CB SER A 444 −21.713 −8.096 −8.936 1.00 51.21 ATOM3397 OG SER A 444 −21.738 −7.735 −7.563 1.00 52.90 ATOM 3398 C SER A 444−19.291 −7.452 −9.165 1.00 51.89 ATOM 3399 O SER A 444 −18.743 −6.610−8.433 1.00 52.46 ATOM 3400 N SER A 445 −18.700 −8.588 −9.558 1.00 52.45ATOM 3401 CA SER A 445 −17.400 −9.030 −9.029 1.00 52.98 ATOM 3402 CB SERA 445 −17.138 −10.497 −9.385 1.00 53.12 ATOM 3403 OG SER A 445 −16.901−10.650 −10.775 1.00 54.45 ATOM 3404 C SER A 445 −16.202 −8.159 −9.4491.00 52.99 ATOM 3405 O SER A 445 −15.184 −8.112 −8.738 1.00 53.28 ATOM3406 N ALA A 446 −16.328 −7.472 −10.588 1.00 52.67 ATOM 3407 CA ALA A446 −15.301 −6.536 −11.063 1.00 52.20 ATOM 3408 CB ALA A 446 −15.695−5.960 −12.425 1.00 52.45 ATOM 3409 C ALA A 446 −14.996 −5.406 −10.0591.00 51.92 ATOM 3410 O ALA A 446 −14.144 −4.553 −10.322 1.00 51.77 ATOM3411 N SER A 447 −15.696 −5.413 −8.919 1.00 51.23 ATOM 3412 CA SER A 447−15.471 −4.453 −7.827 1.00 50.73 ATOM 3413 CB SER A 447 −16.769 −3.706−7.495 1.00 50.70 ATOM 3414 OG SER A 447 −17.765 −4.605 −7.021 1.0050.91 ATOM 3415 C SER A 447 −14.886 −5.072 −6.537 1.00 50.31 ATOM 3416 OSER A 447 −14.604 −4.345 −5.580 1.00 50.16 ATOM 3417 N THR A 448 −14.704−6.394 −6.508 1.00 49.55 ATOM 3418 CA THR A 448 −14.165 −7.070 −5.3141.00 49.05 ATOM 3419 CB THR A 448 −14.579 −8.561 −5.233 1.00 49.15 ATOM3420 OG1 THR A 448 −14.076 −9.255 −6.378 1.00 49.97 ATOM 3421 CG2 THR A448 −16.096 −8.705 −5.180 1.00 49.11 ATOM 3422 C THR A 448 −12.641−6.958 −5.215 1.00 48.34 ATOM 3423 O THR A 448 −11.911 −7.325 −6.1351.00 47.84 ATOM 3424 N ILE A 449 −12.174 −6.444 −4.084 1.00 47.93 ATOM3425 CA ILE A 449 −10.760 −6.150 −3.891 1.00 47.42 ATOM 3426 CB ILE A449 −10.577 −4.798 −3.142 1.00 47.53 ATOM 3427 CG1 ILE A 449 −11.346−3.680 −3.863 1.00 46.60 ATOM 3428 CD1 ILE A 449 −11.727 −2.523 −2.9811.00 45.49 ATOM 3429 CG2 ILE A 449 −9.097 −4.438 −2.999 1.00 46.84 ATOM3430 C ILE A 449 −10.104 −7.299 −3.124 1.00 47.45 ATOM 3431 O ILE A 449−10.606 −7.688 −2.067 1.00 47.66 ATOM 3432 N PRO A 450 −8.993 −7.857−3.663 1.00 47.28 ATOM 3433 CA PRO A 450 −8.202 −8.926 −3.036 1.00 47.25ATOM 3434 CB PRO A 450 −6.982 −9.040 −3.946 1.00 47.03 ATOM 3435 CG PROA 450 −7.430 −8.535 −5.237 1.00 47.02 ATOM 3436 CD PRO A 450 −8.431−7.470 −4.969 1.00 47.07 ATOM 3437 C PRO A 450 −7.731 −8.562 −1.639 1.0047.48 ATOM 3438 O PRO A 450 −7.608 −7.377 −1.322 1.00 47.43 ATOM 3439 NSER A 451 −7.452 −9.587 −0.832 1.00 47.94 ATOM 3440 CA SER A 451 −7.020−9.436 0.568 1.00 48.19 ATOM 3441 CB SER A 451 −7.017 −10.801 1.277 1.0048.52 ATOM 3442 OG SER A 451 −8.297 −11.414 1.235 1.00 49.64 ATOM 3443 CSER A 451 −5.641 −8.799 0.701 1.00 47.83 ATOM 3444 O SER A 451 −5.415−7.963 1.575 1.00 48.39 ATOM 3445 N THR A 452 −4.715 −9.212 −0.158 1.0047.14 ATOM 3446 CA THR A 452 −3.379 −8.613 −0.211 1.00 46.42 ATOM 3447CB THR A 452 −2.323 −9.540 0.434 1.00 46.66 ATOM 3448 OG1 THR A 452−2.518 −10.887 −0.032 1.00 48.38 ATOM 3449 CG2 THR A 452 −2.446 −9.5141.962 1.00 47.77 ATOM 3450 C THR A 452 −3.011 −8.323 −1.673 1.00 44.81ATOM 3451 O THR A 452 −3.348 −9.107 −2.558 1.00 44.97 ATOM 3452 N CYS A453 −2.363 −7.187 −1.931 1.00 43.37 ATOM 3453 CA CYS A 453 −1.971 −6.854−3.306 1.00 41.40 ATOM 3454 CB CYS A 453 −1.918 −5.339 −3.574 1.00 41.01ATOM 3455 SG CYS A 453 −3.187 −4.199 −2.908 1.00 40.58 ATOM 3456 C CYS A453 −0.591 −7.408 −3.602 1.00 40.33 ATOM 3457 O CYS A 453 0.293 −7.370−2.753 1.00 39.88 ATOM 3458 N SER A 454 −0.405 −7.911 −4.812 1.00 39.25ATOM 3459 CA SER A 454 0.937 −8.142 −5.336 1.00 38.68 ATOM 3460 CB SER A454 1.222 −9.638 −5.484 1.00 38.56 ATOM 3461 OG SER A 454 0.276 −10.251−6.349 1.00 40.44 ATOM 3462 C SER A 454 1.047 −7.450 −6.690 1.00 37.78ATOM 3463 O SER A 454 0.030 −7.187 −7.347 1.00 36.71 ATOM 3464 N GLY A455 2.275 −7.175 −7.111 1.00 37.06 ATOM 3465 CA GLY A 455 2.514 −6.613−8.431 1.00 36.79 ATOM 3466 C GLY A 455 2.493 −7.658 −9.539 1.00 36.18ATOM 3467 O GLY A 455 3.410 −7.708 −10.367 1.00 36.73 ATOM 3468 N ALA A456 1.445 −8.480 −9.562 1.00 35.26 ATOM 3469 CA ALA A 456 1.321 −9.584−10.512 1.00 34.46 ATOM 3470 CB ALA A 456 0.125 −10.454 −10.158 1.0034.50 ATOM 3471 C ALA A 456 1.195 −9.111 −11.963 1.00 34.16 ATOM 3472 OALA A 456 0.283 −8.353 −12.301 1.00 33.56 ATOM 3473 N SER A 457 2.097−9.588 −12.817 1.00 33.11 ATOM 3474 CA SER A 457 2.013 −9.302 −14.2411.00 32.60 ATOM 3475 CB SER A 457 3.022 −8.219 −14.635 1.00 32.50 ATOM3476 OG SER A 457 4.352 −8.691 −14.519 1.00 33.28 ATOM 3477 C SER A 4572.228 −10.575 −15.044 1.00 32.31 ATOM 3478 O SER A 457 2.641 −11.605−14.494 1.00 32.32 ATOM 3479 N VAL A 458 1.908 −10.511 −16.330 1.0031.12 ATOM 3480 CA VAL A 458 2.063 −11.629 −17.246 1.00 31.09 ATOM 3481CB VAL A 458 0.682 −12.199 −17.659 1.00 31.00 ATOM 3482 CG1 VAL A 4580.806 −13.173 −18.830 1.00 30.72 ATOM 3483 CG2 VAL A 458 −0.014 −12.847−16.459 1.00 31.08 ATOM 3484 C VAL A 458 2.817 −11.144 −18.480 1.0030.88 ATOM 3485 O VAL A 458 2.401 −10.177 −19.126 1.00 29.70 ATOM 3486 NVAL A 459 3.924 −11.811 −18.805 1.00 30.81 ATOM 3487 CA VAL A 459 4.643−11.525 −20.051 1.00 30.95 ATOM 3488 CB VAL A 459 6.046 −12.172 −20.0711.00 31.34 ATOM 3489 CG1 VAL A 459 6.664 −12.102 −21.492 1.00 30.45 ATOM3490 CG2 VAL A 459 6.947 −11.522 −19.030 1.00 32.03 ATOM 3491 C VAL A459 3.805 −12.032 −21.227 1.00 31.37 ATOM 3492 O VAL A 459 3.443 −13.214−21.288 1.00 31.84 ATOM 3493 N GLY A 460 3.480 −11.137 −22.154 1.0030.71 ATOM 3494 CA GLY A 460 2.596 −11.495 −23.258 1.00 30.77 ATOM 3495C GLY A 460 3.349 −11.799 −24.536 1.00 30.66 ATOM 3496 O GLY A 460 4.585−11.773 −24.582 1.00 30.90 ATOM 3497 N SER A 461 2.606 −12.094 −25.5841.00 30.44 ATOM 3498 CA SER A 461 3.219 −12.227 −26.877 1.00 31.06 ATOM3499 CB SER A 461 3.301 −13.695 −27.308 1.00 31.44 ATOM 3500 OG SER A461 2.018 −14.278 −27.419 1.00 34.77 ATOM 3501 C SER A 461 2.463 −11.357−27.864 1.00 30.21 ATOM 3502 O SER A 461 1.246 −11.156 −27.736 1.0030.87 ATOM 3503 N TYR A 462 3.192 −10.822 −28.836 1.00 28.84 ATOM 3504CA TYR A 462 2.651 −9.797 −29.712 1.00 28.13 ATOM 3505 CB TYR A 4623.365 −8.471 −29.426 1.00 26.74 ATOM 3506 CG TYR A 462 3.264 −8.098−27.976 1.00 25.28 ATOM 3507 CD1 TYR A 462 2.184 −7.335 −27.508 1.0023.75 ATOM 3508 CE1 TYR A 462 2.066 −7.022 −26.162 1.00 23.41 ATOM 3509CZ TYR A 462 3.030 −7.458 −25.268 1.00 23.32 ATOM 3510 OH TYR A 4622.907 −7.155 −23.941 1.00 23.50 ATOM 3511 CE2 TYR A 462 4.113 −8.226−25.698 1.00 22.57 ATOM 3512 CD2 TYR A 462 4.223 −8.537 −27.050 1.0024.71 ATOM 3513 C TYR A 462 2.844 −10.191 −31.152 1.00 28.83 ATOM 3514 OTYR A 462 3.898 −10.697 −31.529 1.00 29.33 ATOM 3515 N SER A 463 1.828−9.973 −31.961 1.00 29.66 ATOM 3516 CA SER A 463 1.970 −10.202 −33.3881.00 30.77 ATOM 3517 CB SER A 463 1.424 −11.574 −33.784 1.00 30.99 ATOM3518 OG SER A 463 0.168 −11.815 −33.192 1.00 33.22 ATOM 3519 C SER A 4631.311 −9.082 −34.170 1.00 31.28 ATOM 3520 O SER A 463 0.329 −8.481−33.723 1.00 30.40 ATOM 3521 N ARG A 464 1.886 −8.789 −35.330 1.00 31.94ATOM 3522 CA ARG A 464 1.377 −7.775 −36.225 1.00 32.97 ATOM 3523 CB ARGA 464 2.362 −7.620 −37.385 1.00 33.68 ATOM 3524 CG ARG A 464 2.353−6.274 −38.061 1.00 36.94 ATOM 3525 CD ARG A 464 3.502 −6.206 −39.0881.00 42.45 ATOM 3526 NE ARG A 464 4.794 −6.065 −38.415 1.00 45.53 ATOM3527 CZ ARG A 464 5.416 −4.903 −38.227 1.00 47.50 ATOM 3528 NH1 ARG A464 4.882 −3.775 −38.688 1.00 49.50 ATOM 3529 NH2 ARG A 464 6.580 −4.863−37.592 1.00 48.59 ATOM 3530 C ARG A 464 −0.017 −8.171 −36.741 1.0033.03 ATOM 3531 O ARG A 464 −0.166 −9.228 −37.358 1.00 33.00 ATOM 3532 NPRO A 465 −1.053 −7.333 −36.479 1.00 32.72 ATOM 3533 CA PRO A 465 −2.344−7.593 −37.131 1.00 32.69 ATOM 3534 CB PRO A 465 −3.274 −6.504 −36.5581.00 32.50 ATOM 3535 CG PRO A 465 −2.581 −5.981 −35.345 1.00 32.47 ATOM3536 CD PRO A 465 −1.102 −6.134 −35.618 1.00 32.83 ATOM 3537 C PRO A 465−2.189 −7.421 −38.642 1.00 33.30 ATOM 3538 O PRO A 465 −1.332 −6.661−39.097 1.00 33.20 ATOM 3539 N THR A 466 −2.990 −8.136 −39.412 1.0034.26 ATOM 3540 CA THR A 466 −2.810 −8.131 −40.855 1.00 35.62 ATOM 3541CB THR A 466 −2.264 −9.486 −41.370 1.00 35.33 ATOM 3542 OG1 THR A 466−3.225 −10.512 −41.136 1.00 36.81 ATOM 3543 CG2 THR A 466 −0.965 −9.848−40.656 1.00 35.91 ATOM 3544 C THR A 466 −4.076 −7.711 −41.600 1.0036.15 ATOM 3545 O THR A 466 −3.983 −7.077 −42.648 1.00 36.93 ATOM 3546 NALA A 467 −5.242 −8.051 −41.048 1.00 36.82 ATOM 3547 CA ALA A 467 −6.540−7.648 −41.609 1.00 37.30 ATOM 3548 CB ALA A 467 −7.663 −8.403 −40.9301.00 37.04 ATOM 3549 C ALA A 467 −6.767 −6.136 −41.509 1.00 38.13 ATOM3550 O ALA A 467 −6.715 −5.556 −40.417 1.00 37.96 ATOM 3551 N THR A 468−7.011 −5.502 −42.653 1.00 38.66 ATOM 3552 CA THR A 468 −7.146 −4.050−42.702 1.00 39.18 ATOM 3553 CB THR A 468 −5.970 −3.406 −43.428 1.0039.43 ATOM 3554 OG1 THR A 468 −5.955 −3.879 −44.778 1.00 40.06 ATOM 3555CG2 THR A 468 −4.637 −3.717 −42.734 1.00 39.67 ATOM 3556 C THR A 468−8.405 −3.591 −43.427 1.00 39.33 ATOM 3557 O THR A 468 −8.468 −2.451−43.890 1.00 39.79 ATOM 3558 N SER A 469 −9.403 −4.457 −43.529 1.0039.12 ATOM 3559 CA SER A 469 −10.651 −4.065 −44.176 1.00 39.41 ATOM 3560CB SER A 469 −10.624 −4.369 −45.684 1.00 39.77 ATOM 3561 OG SER A 469−10.476 −5.763 −45.916 1.00 41.02 ATOM 3562 C SER A 469 −11.850 −4.732−43.537 1.00 38.54 ATOM 3563 O SER A 469 −11.771 −5.856 −43.046 1.0038.85 ATOM 3564 N PHE A 470 −12.962 −4.014 −43.558 1.00 38.00 ATOM 3565CA PHE A 470 −14.216 −4.507 −43.031 1.00 37.29 ATOM 3566 CB PHE A 470−14.880 −3.406 −42.220 1.00 36.87 ATOM 3567 CG PHE A 470 −14.277 −3.212−40.865 1.00 35.43 ATOM 3568 CD1 PHE A 470 −13.146 −2.428 −40.696 1.0035.15 ATOM 3569 CE1 PHE A 470 −12.589 −2.252 −39.437 1.00 33.73 ATOM3570 CZ PHE A 470 −13.159 −2.852 −38.332 1.00 35.04 ATOM 3571 CE2 PHE A470 −14.292 −3.638 −38.479 1.00 36.26 ATOM 3572 CD2 PHE A 470 −14.844−3.816 −39.751 1.00 35.71 ATOM 3573 C PHE A 470 −15.128 −4.923 −44.1811.00 37.34 ATOM 3574 O PHE A 470 −15.059 −4.337 −45.258 1.00 37.21 ATOM3575 N PRO A 471 −15.987 −5.931 −43.959 1.00 37.46 ATOM 3576 CA PRO A471 −16.983 −6.243 −44.985 1.00 38.04 ATOM 3577 CB PRO A 471 −17.790−7.383 −44.361 1.00 37.60 ATOM 3578 CG PRO A 471 −16.877 −7.986 −43.3371.00 38.09 ATOM 3579 CD PRO A 471 −16.093 −6.828 −42.795 1.00 37.31 ATOM3580 C PRO A 471 −17.879 −5.033 −45.231 1.00 38.96 ATOM 3581 O PRO A 471−18.108 −4.245 −44.306 1.00 38.78 ATOM 3582 N PRO A 472 −18.378 −4.869−46.471 1.00 39.78 ATOM 3583 CA PRO A 472 −19.238 −3.723 −46.771 1.0039.87 ATOM 3584 CB PRO A 472 −19.378 −3.780 −48.293 1.00 40.09 ATOM 3585CG PRO A 472 −19.171 −5.225 −48.635 1.00 40.28 ATOM 3586 CD PRO A 472−18.171 −5.740 −47.649 1.00 40.02 ATOM 3587 C PRO A 472 −20.604 −3.864−46.114 1.00 39.75 ATOM 3588 O PRO A 472 −21.017 −4.980 −45.798 1.0040.10 ATOM 3589 N SER A 473 −21.268 −2.734 −45.881 1.00 39.71 ATOM 3590CA SER A 473 −22.675 −2.696 −45.467 1.00 39.61 ATOM 3591 CB SER A 473−23.571 −3.070 −46.651 1.00 40.16 ATOM 3592 OG SER A 473 −23.468 −2.074−47.658 1.00 42.55 ATOM 3593 C SER A 473 −23.043 −3.509 −44.221 1.0038.94 ATOM 3594 O SER A 473 −24.041 −4.258 −44.210 1.00 39.07 ATOM 3595N GLN A 474 −22.257 −3.340 −43.159 1.00 37.75 ATOM 3596 CA GLN A 474−22.558 −3.993 −41.888 1.00 36.60 ATOM 3597 CB GLN A 474 −21.291 −4.205−41.057 1.00 36.41 ATOM 3598 CG GLN A 474 −20.331 −5.169 −41.732 1.0036.24 ATOM 3599 CD GLN A 474 −19.295 −5.757 −40.795 1.00 36.45 ATOM 3600OE1 GLN A 474 −18.478 −5.040 −40.211 1.00 35.30 ATOM 3601 NE2 GLN A 474−19.304 −7.077 −40.671 1.00 36.85 ATOM 3602 C GLN A 474 −23.620 −3.191−41.149 1.00 36.40 ATOM 3603 O GLN A 474 −23.329 −2.444 −40.208 1.0035.99 ATOM 3604 N THR A 475 −24.859 −3.350 −41.616 1.00 35.59 ATOM 3605CA THR A 475 −26.012 −2.595 −41.148 1.00 35.20 ATOM 3606 CB THR A 475−27.051 −2.478 −42.287 1.00 35.73 ATOM 3607 OG1 THR A 475 −27.120 −3.737−42.959 1.00 36.85 ATOM 3608 CG2 THR A 475 −26.642 −1.418 −43.310 1.0034.51 ATOM 3609 C THR A 475 −26.635 −3.256 −39.910 1.00 35.16 ATOM 3610O THR A 475 −26.363 −4.420 −39.622 1.00 34.47 ATOM 3611 N PRO A 476−27.453 −2.510 −39.148 1.00 35.36 ATOM 3612 CA PRO A 476 −27.990 −3.111−37.923 1.00 36.40 ATOM 3613 CB PRO A 476 −28.567 −1.910 −37.167 1.0036.04 ATOM 3614 CG PRO A 476 −28.890 −0.912 −38.230 1.00 35.98 ATOM 3615CD PRO A 476 −27.907 −1.119 −39.339 1.00 35.70 ATOM 3616 C PRO A 476−29.085 −4.171 −38.158 1.00 37.45 ATOM 3617 O PRO A 476 −29.654 −4.244−39.254 1.00 36.95 ATOM 3618 N LYS A 477 −29.342 −4.984 −37.133 1.0039.01 ATOM 3619 CA LYS A 477 −30.472 −5.911 −37.111 1.00 41.20 ATOM 3620CB LYS A 477 −30.541 −6.641 −35.774 1.00 41.46 ATOM 3621 CG LYS A 477−29.665 −7.850 −35.604 1.00 41.87 ATOM 3622 CD LYS A 477 −29.939 −8.517−34.237 1.00 42.36 ATOM 3623 CE LYS A 477 −29.996 −7.497 −33.076 1.0044.55 ATOM 3624 NZ LYS A 477 −29.705 −8.110 −31.718 1.00 44.56 ATOM 3625C LYS A 477 −31.766 −5.118 −37.230 1.00 42.27 ATOM 3626 O LYS A 477−31.818 −3.960 −36.798 1.00 42.18 ATOM 3627 N PRO A 478 −32.831 −5.743−37.780 1.00 43.46 ATOM 3628 CA PRO A 478 −34.150 −5.106 −37.669 1.0044.02 ATOM 3629 CB PRO A 478 −35.106 −6.144 −38.267 1.00 43.89 ATOM 3630CG PRO A 478 −34.255 −7.033 −39.105 1.00 44.08 ATOM 3631 CD PRO A 478−32.885 −7.034 −38.493 1.00 43.41 ATOM 3632 C PRO A 478 −34.480 −4.892−36.194 1.00 44.52 ATOM 3633 O PRO A 478 −34.197 −5.769 −35.364 1.0044.73 ATOM 3634 N GLY A 479 −35.043 −3.728 −35.874 1.00 45.20 ATOM 3635CA GLY A 479 −35.421 −3.395 −34.494 1.00 45.90 ATOM 3636 C GLY A 479−34.386 −2.601 −33.711 1.00 46.37 ATOM 3637 O GLY A 479 −34.576 −2.331−32.520 1.00 46.98 ATOM 3638 N VAL A 480 −33.282 −2.244 −34.367 1.0046.28 ATOM 3639 CA VAL A 480 −32.261 −1.383 −33.760 1.00 46.15 ATOM 3640CB VAL A 480 −30.820 −1.863 −34.121 1.00 46.20 ATOM 3641 CG1 VAL A 480−29.755 −0.899 −33.584 1.00 45.85 ATOM 3642 CG2 VAL A 480 −30.569 −3.281−33.603 1.00 46.25 ATOM 3643 C VAL A 480 −32.498 0.046 −34.260 1.0045.93 ATOM 3644 O VAL A 480 −32.673 0.240 −35.465 1.00 46.15 ATOM 3645 NPRO A 481 −32.534 1.049 −33.344 1.00 45.70 ATOM 3646 CA PRO A 481−32.648 2.443 −33.804 1.00 45.36 ATOM 3647 CB PRO A 481 −32.388 3.266−32.542 1.00 45.39 ATOM 3648 CG PRO A 481 −32.778 2.375 −31.427 1.0045.52 ATOM 3649 CD PRO A 481 −32.481 0.962 −31.873 1.00 45.83 ATOM 3650C PRO A 481 −31.609 2.762 −34.877 1.00 45.08 ATOM 3651 O PRO A 481−30.405 2.555 −34.681 1.00 44.45 ATOM 3652 N SER A 482 −32.100 3.241−36.011 1.00 44.83 ATOM 3653 CA SER A 482 −31.281 3.485 −37.180 1.0044.52 ATOM 3654 CB SER A 482 −31.502 2.375 −38.211 1.00 44.74 ATOM 3655OG SER A 482 −30.769 2.622 −39.399 1.00 45.89 ATOM 3656 C SER A 482−31.661 4.836 −37.765 1.00 43.84 ATOM 3657 O SER A 482 −32.836 5.219−37.741 1.00 44.07 ATOM 3658 N GLY A 483 −30.667 5.550 −38.282 1.0042.68 ATOM 3659 CA GLY A 483 −30.872 6.872 −38.853 1.00 41.64 ATOM 3660C GLY A 483 −30.085 7.095 −40.130 1.00 41.05 ATOM 3661 O GLY A 483−29.430 6.179 −40.647 1.00 41.09 ATOM 3662 N THR A 484 −30.155 8.317−40.647 1.00 40.22 ATOM 3663 CA THR A 484 −29.461 8.677 −41.888 1.0039.91 ATOM 3664 CB THR A 484 −30.148 9.876 −42.619 1.00 40.21 ATOM 3665OG1 THR A 484 −30.115 11.040 −41.780 1.00 41.48 ATOM 3666 CG2 THR A 484−31.604 9.541 −43.000 1.00 40.86 ATOM 3667 C THR A 484 −27.995 9.033−41.603 1.00 38.43 ATOM 3668 O THR A 484 −27.669 9.421 −40.483 1.0038.52 ATOM 3669 N PRO A 485 −27.109 8.893 −42.612 1.00 37.36 ATOM 3670CA PRO A 485 −25.695 9.226 −42.413 1.00 36.29 ATOM 3671 CB PRO A 485−25.077 8.974 −43.795 1.00 36.75 ATOM 3672 CG PRO A 485 −25.997 7.985−44.442 1.00 37.04 ATOM 3673 CD PRO A 485 −27.359 8.393 −43.976 1.0037.30 ATOM 3674 C PRO A 485 −25.460 10.684 −41.988 1.00 35.20 ATOM 3675O PRO A 485 −26.201 11.599 −42.396 1.00 34.39 ATOM 3676 N TYR A 486−24.428 10.887 −41.174 1.00 33.60 ATOM 3677 CA TYR A 486 −24.025 12.233−40.782 1.00 32.74 ATOM 3678 CB TYR A 486 −22.821 12.180 −39.826 1.0032.31 ATOM 3679 CG TYR A 486 −22.348 13.564 −39.452 1.00 32.79 ATOM 3680CD1 TYR A 486 −21.243 14.146 −40.083 1.00 31.74 ATOM 3681 CE1 TYR A 486−20.827 15.430 −39.742 1.00 30.85 ATOM 3682 CZ TYR A 486 −21.527 16.141−38.778 1.00 31.99 ATOM 3683 OH TYR A 486 −21.143 17.423 −38.427 1.0032.20 ATOM 3684 CE2 TYR A 486 −22.629 15.588 −38.160 1.00 31.88 ATOM3685 CD2 TYR A 486 −23.036 14.311 −38.500 1.00 32.49 ATOM 3686 C TYR A486 −23.652 13.082 −41.999 1.00 31.99 ATOM 3687 O TYR A 486 −22.94912.602 −42.900 1.00 31.62 ATOM 3688 N THR A 487 −24.106 14.336 −42.0041.00 31.14 ATOM 3689 CA THR A 487 −23.676 15.336 −42.986 1.00 31.43 ATOM3690 CB THR A 487 −24.879 15.785 −43.869 1.00 31.94 ATOM 3691 OG1 THR A487 −25.321 14.665 −44.644 1.00 35.19 ATOM 3692 CG2 THR A 487 −24.48916.904 −44.810 1.00 32.95 ATOM 3693 C THR A 487 −23.110 16.561 −42.2611.00 29.79 ATOM 3694 O THR A 487 −23.761 17.078 −41.363 1.00 29.20 ATOM3695 N PRO A 488 −21.901 17.027 −42.644 1.00 29.20 ATOM 3696 CA PRO A488 −21.309 18.228 −42.005 1.00 28.77 ATOM 3697 CB PRO A 488 −19.98818.435 −42.763 1.00 28.75 ATOM 3698 CG PRO A 488 −19.684 17.126 −43.4081.00 29.52 ATOM 3699 CD PRO A 488 −21.010 16.448 −43.667 1.00 29.41 ATOM3700 C PRO A 488 −22.175 19.463 −42.194 1.00 28.44 ATOM 3701 O PRO A 488−23.003 19.499 −43.116 1.00 28.51 ATOM 3702 N LEU A 489 −21.971 20.469−41.345 1.00 27.28 ATOM 3703 CA LEU A 489 −22.606 21.775 −41.522 1.0026.52 ATOM 3704 CB LEU A 489 −22.269 22.708 −40.365 1.00 27.19 ATOM 3705CG LEU A 489 −22.805 22.303 −38.987 1.00 27.53 ATOM 3706 CD1 LEU A 489−22.233 23.242 −37.929 1.00 26.81 ATOM 3707 CD2 LEU A 489 −24.332 22.323−38.970 1.00 28.98 ATOM 3708 C LEU A 489 −22.137 22.402 −42.833 1.0026.65 ATOM 3709 O LEU A 489 −20.983 22.210 −43.245 1.00 25.40 ATOM 3710N PRO A 490 −23.030 23.153 −43.503 1.00 26.84 ATOM 3711 CA PRO A 490−22.636 23.745 −44.786 1.00 26.44 ATOM 3712 CB PRO A 490 −23.983 24.107−45.432 1.00 27.00 ATOM 3713 CG PRO A 490 −24.900 24.341 −44.289 1.0027.68 ATOM 3714 CD PRO A 490 −24.425 23.475 −43.137 1.00 26.91 ATOM 3715C PRO A 490 −21.737 24.982 −44.668 1.00 26.22 ATOM 3716 O PRO A 490−21.826 25.729 −43.698 1.00 25.74 ATOM 3717 N CYS A 491 −20.858 25.182−45.650 1.00 26.06 ATOM 3718 CA CYS A 491 −20.079 26.412 −45.754 1.0026.88 ATOM 3719 CB CYS A 491 −18.630 26.194 −45.302 1.00 27.00 ATOM 3720SG CYS A 491 −18.450 25.196 −43.819 1.00 27.23 ATOM 3721 C CYS A 491−20.032 26.822 −47.217 1.00 27.27 ATOM 3722 O CYS A 491 −20.369 26.026−48.083 1.00 27.34 ATOM 3723 N ALA A 492 −19.577 28.045 −47.484 1.0028.05 ATOM 3724 CA ALA A 492 −19.205 28.449 −48.845 1.00 29.40 ATOM 3725CB ALA A 492 −18.837 29.928 −48.866 1.00 29.49 ATOM 3726 C ALA A 492−18.023 27.599 −49.320 1.00 30.37 ATOM 3727 O ALA A 492 −17.310 26.998−48.497 1.00 30.36 ATOM 3728 N THR A 493 −17.828 27.508 −50.633 1.0031.27 ATOM 3729 CA THR A 493 −16.612 26.883 −51.163 1.00 32.43 ATOM 3730CB THR A 493 −16.845 26.234 −52.533 1.00 33.22 ATOM 3731 OG1 THR A 493−17.944 25.324 −52.431 1.00 38.55 ATOM 3732 CG2 THR A 493 −15.590 25.464−52.996 1.00 33.71 ATOM 3733 C THR A 493 −15.596 28.006 −51.254 1.0031.35 ATOM 3734 O THR A 493 −15.916 29.068 −51.795 1.00 31.58 ATOM 3735N PRO A 494 −14.390 27.815 −50.682 1.00 30.44 ATOM 3736 CA PRO A 494−13.464 28.947 −50.696 1.00 30.01 ATOM 3737 CB PRO A 494 −12.414 28.555−49.645 1.00 30.26 ATOM 3738 CG PRO A 494 −12.416 27.077 −49.658 1.0030.56 ATOM 3739 CD PRO A 494 −13.815 26.635 −49.997 1.00 30.85 ATOM 3740C PRO A 494 −12.809 29.089 −52.060 1.00 28.81 ATOM 3741 O PRO A 494−12.801 28.137 −52.834 1.00 28.82 ATOM 3742 N THR A 495 −12.260 30.258−52.352 1.00 28.43 ATOM 3743 CA THR A 495 −11.551 30.419 −53.623 1.0028.44 ATOM 3744 CB THR A 495 −11.885 31.748 −54.319 1.00 28.68 ATOM 3745OG1 THR A 495 −11.449 32.839 −53.500 1.00 30.39 ATOM 3746 CG2 THR A 495−13.383 31.858 −54.564 1.00 29.85 ATOM 3747 C THR A 495 −10.057 30.335−53.404 1.00 27.94 ATOM 3748 O THR A 495 −9.289 30.159 −54.352 1.0027.63 ATOM 3749 N SER A 496 −9.671 30.463 −52.139 1.00 27.51 ATOM 3750CA SER A 496 −8.279 30.492 −51.722 1.00 27.18 ATOM 3751 CB SER A 496−7.928 31.916 −51.329 1.00 27.72 ATOM 3752 OG SER A 496 −6.531 32.076−51.240 1.00 32.60 ATOM 3753 C SER A 496 −8.134 29.583 −50.501 1.0025.85 ATOM 3754 O SER A 496 −9.024 29.548 −49.634 1.00 25.10 ATOM 3755 NVAL A 497 −7.039 28.824 −50.430 1.00 24.43 ATOM 3756 CA VAL A 497 −6.80128.032 −49.209 1.00 22.17 ATOM 3757 CB VAL A 497 −7.281 26.511 −49.2901.00 23.00 ATOM 3758 CG1 VAL A 497 −6.224 25.445 −48.881 1.00 22.48 ATOM3759 CG2 VAL A 497 −8.049 26.161 −50.578 1.00 22.62 ATOM 3760 C VAL A497 −5.388 28.251 −48.672 1.00 21.13 ATOM 3761 O VAL A 497 −4.419 28.359−49.439 1.00 20.38 ATOM 3762 N ALA A 498 −5.302 28.395 −47.355 1.0019.70 ATOM 3763 CA ALA A 498 −4.020 28.576 −46.702 1.00 18.92 ATOM 3764CB ALA A 498 −4.226 29.126 −45.290 1.00 19.31 ATOM 3765 C ALA A 498−3.396 27.185 −46.655 1.00 18.67 ATOM 3766 O ALA A 498 −3.966 26.266−46.047 1.00 19.29 ATOM 3767 N VAL A 499 −2.252 27.021 −47.319 1.0017.23 ATOM 3768 CA VAL A 499 −1.551 25.735 −47.361 1.00 16.09 ATOM 3769CB VAL A 499 −1.165 25.347 −48.814 1.00 16.97 ATOM 3770 CG1 VAL A 499−0.403 23.984 −48.863 1.00 16.08 ATOM 3771 CG2 VAL A 499 −2.413 25.291−49.696 1.00 17.03 ATOM 3772 C VAL A 499 −0.306 25.841 −46.491 1.0015.95 ATOM 3773 O VAL A 499 0.604 26.607 −46.791 1.00 16.15 ATOM 3774 NTHR A 500 −0.279 25.085 −45.404 1.00 15.16 ATOM 3775 CA THR A 500 0.86325.116 −44.505 1.00 14.85 ATOM 3776 CB THR A 500 0.415 24.916 −43.0351.00 14.76 ATOM 3777 OG1 THR A 500 −0.403 26.022 −42.635 1.00 16.00 ATOM3778 CG2 THR A 500 1.639 24.856 −42.136 1.00 15.39 ATOM 3779 C THR A 5001.796 23.993 −44.932 1.00 14.99 ATOM 3780 O THR A 500 1.480 22.804−44.792 1.00 14.81 ATOM 3781 N PHE A 501 2.941 24.370 −45.481 1.00 14.71ATOM 3782 CA PHE A 501 3.981 23.411 −45.793 1.00 15.64 ATOM 3783 CB PHEA 501 4.943 23.964 −46.832 1.00 15.86 ATOM 3784 CG PHE A 501 4.28924.172 −48.168 1.00 18.38 ATOM 3785 CD1 PHE A 501 3.676 25.388 −48.4691.00 19.85 ATOM 3786 CE1 PHE A 501 3.052 25.581 −49.709 1.00 21.58 ATOM3787 CZ PHE A 501 3.015 24.547 −50.642 1.00 19.80 ATOM 3788 CE2 PHE A501 3.607 23.324 −50.356 1.00 21.69 ATOM 3789 CD2 PHE A 501 4.231 23.134−49.095 1.00 21.96 ATOM 3790 C PHE A 501 4.711 23.009 −44.536 1.00 15.81ATOM 3791 O PHE A 501 5.207 23.852 −43.804 1.00 16.78 ATOM 3792 N HIS A502 4.789 21.698 −44.317 1.00 14.85 ATOM 3793 CA HIS A 502 5.239 21.175−43.027 1.00 14.17 ATOM 3794 CB HIS A 502 3.987 20.565 −42.356 1.0014.85 ATOM 3795 CG HIS A 502 4.221 19.875 −41.054 1.00 13.69 ATOM 3796ND1 HIS A 502 4.819 18.637 −40.966 1.00 12.55 ATOM 3797 CE1 HIS A 5024.816 18.241 −39.702 1.00 15.76 ATOM 3798 NE2 HIS A 502 4.191 19.155−38.980 1.00 14.42 ATOM 3799 CD2 HIS A 502 3.797 20.183 −39.804 1.0014.92 ATOM 3800 C HIS A 502 6.317 20.161 −43.412 1.00 14.16 ATOM 3801 OHIS A 502 6.013 19.043 −43.824 1.00 13.85 ATOM 3802 N GLU A 503 7.57720.590 −43.352 1.00 13.63 ATOM 3803 CA GLU A 503 8.678 19.821 −43.9681.00 14.46 ATOM 3804 CB GLU A 503 9.434 20.712 −44.996 1.00 14.22 ATOM3805 CG GLU A 503 10.782 20.121 −45.524 1.00 16.31 ATOM 3806 CD GLU A503 10.620 18.973 −46.539 1.00 21.32 ATOM 3807 OE1 GLU A 503 11.52318.819 −47.393 1.00 21.21 ATOM 3808 OE2 GLU A 503 9.609 18.230 −46.5101.00 20.10 ATOM 3809 C GLU A 503 9.657 19.322 −42.917 1.00 14.22 ATOM3810 O GLU A 503 10.175 20.121 −42.131 1.00 15.31 ATOM 3811 N LEU A 5049.960 18.027 −42.927 1.00 14.92 ATOM 3812 CA LEU A 504 11.026 17.518−42.052 1.00 15.49 ATOM 3813 CB LEU A 504 10.658 16.147 −41.489 1.0016.33 ATOM 3814 CG LEU A 504 9.479 16.178 −40.498 1.00 17.19 ATOM 3815CD1 LEU A 504 8.922 14.753 −40.320 1.00 19.08 ATOM 3816 CD2 LEU A 5049.953 16.723 −39.198 1.00 17.28 ATOM 3817 C LEU A 504 12.318 17.428−42.846 1.00 16.59 ATOM 3818 O LEU A 504 12.403 16.656 −43.785 1.0016.72 ATOM 3819 N VAL A 505 13.317 18.201 −42.444 1.00 17.20 ATOM 3820CA VAL A 505 14.592 18.235 −43.154 1.00 19.16 ATOM 3821 CB VAL A 50514.548 19.141 −44.418 1.00 18.88 ATOM 3822 CG1 VAL A 505 14.028 20.539−44.090 1.00 19.28 ATOM 3823 CG2 VAL A 505 15.948 19.219 −45.095 1.0021.65 ATOM 3824 C VAL A 505 15.674 18.705 −42.188 1.00 20.00 ATOM 3825 OVAL A 505 15.595 19.785 −41.595 1.00 19.92 ATOM 3826 N SER A 506 16.68517.868 −42.011 1.00 21.73 ATOM 3827 CA SER A 506 17.761 18.216 −41.1041.00 23.33 ATOM 3828 CB SER A 506 18.570 16.974 −40.771 1.00 23.74 ATOM3829 OG SER A 506 19.583 17.320 −39.847 1.00 28.30 ATOM 3830 C SER A 50618.646 19.284 −41.759 1.00 23.03 ATOM 3831 O SER A 506 19.070 19.139−42.908 1.00 23.20 ATOM 3832 N THR A 507 18.888 20.371 −41.049 1.0024.01 ATOM 3833 CA THR A 507 19.685 21.464 −41.600 1.00 25.15 ATOM 3834CB THR A 507 18.845 22.725 −41.833 1.00 25.01 ATOM 3835 OG1 THR A 50718.104 23.015 −40.650 1.00 24.43 ATOM 3836 CG2 THR A 507 17.891 22.536−43.000 1.00 24.71 ATOM 3837 C THR A 507 20.795 21.812 −40.623 1.0027.09 ATOM 3838 O THR A 507 20.729 21.448 −39.451 1.00 26.15 ATOM 3839 NGLN A 508 21.798 22.536 −41.113 1.00 29.34 ATOM 3840 CA GLN A 508 22.91222.986 −40.272 1.00 32.54 ATOM 3841 CB GLN A 508 24.239 22.542 −40.8971.00 32.54 ATOM 3842 CG GLN A 508 24.369 21.010 −40.972 1.00 34.32 ATOM3843 CD GLN A 508 25.400 20.515 −41.991 1.00 36.23 ATOM 3844 OE1 GLN A508 26.283 19.700 −41.660 1.00 41.79 ATOM 3845 NE2 GLN A 508 25.27920.977 −43.242 1.00 40.42 ATOM 3846 C GLN A 508 22.827 24.502 −40.1001.00 33.06 ATOM 3847 O GLN A 508 22.136 25.178 −40.873 1.00 32.49 ATOM3848 N PHE A 509 23.494 25.037 −39.075 1.00 33.80 ATOM 3849 CA PHE A 50923.432 26.476 −38.782 1.00 35.03 ATOM 3850 CB PHE A 509 24.413 26.837−37.651 1.00 36.75 ATOM 3851 CG PHE A 509 24.481 28.315 −37.340 1.0039.07 ATOM 3852 CD1 PHE A 509 23.592 28.893 −36.428 1.00 41.58 ATOM 3853CE1 PHE A 509 23.642 30.265 −36.140 1.00 42.61 ATOM 3854 CZ PHE A 50924.603 31.073 −36.766 1.00 41.78 ATOM 3855 CE2 PHE A 509 25.507 30.503−37.678 1.00 42.48 ATOM 3856 CD2 PHE A 509 25.441 29.127 −37.955 1.0041.46 ATOM 3857 C PHE A 509 23.712 27.311 −40.040 1.00 34.58 ATOM 3858 OPHE A 509 24.614 26.990 −40.815 1.00 34.58 ATOM 3859 N GLY A 510 22.91228.355 −40.256 1.00 33.85 ATOM 3860 CA GLY A 510 23.101 29.241 −41.4071.00 33.22 ATOM 3861 C GLY A 510 22.352 28.826 −42.671 1.00 32.36 ATOM3862 O GLY A 510 22.369 29.545 −43.679 1.00 32.97 ATOM 3863 N GLN A 51121.705 27.663 −42.628 1.00 30.50 ATOM 3864 CA GLN A 511 20.885 27.217−43.745 1.00 29.02 ATOM 3865 CB GLN A 511 21.026 25.712 −43.931 1.0028.92 ATOM 3866 CG GLN A 511 22.436 25.276 −44.349 1.00 29.91 ATOM 3867CD GLN A 511 22.571 23.776 −44.439 1.00 31.36 ATOM 3868 OE1 GLN A 51121.760 23.036 −43.879 1.00 31.69 ATOM 3869 NE2 GLN A 511 23.590 23.309−45.160 1.00 30.72 ATOM 3870 C GLN A 511 19.418 27.619 −43.543 1.0027.82 ATOM 3871 O GLN A 511 18.928 27.695 −42.399 1.00 27.36 ATOM 3872 NTHR A 512 18.727 27.895 −44.650 1.00 25.92 ATOM 3873 CA THR A 512 17.30528.271 −44.613 1.00 24.51 ATOM 3874 CB THR A 512 17.126 29.763 −44.9941.00 24.95 ATOM 3875 OG1 THR A 512 17.769 30.580 −44.004 1.00 27.43 ATOM3876 CG2 THR A 512 15.653 30.151 −45.069 1.00 25.94 ATOM 3877 C THR A512 16.536 27.384 −45.600 1.00 23.09 ATOM 3878 O THR A 512 16.994 27.152−46.717 1.00 22.75 ATOM 3879 N VAL A 513 15.376 26.877 −45.200 1.0020.62 ATOM 3880 CA VAL A 513 14.593 26.074 −46.136 1.00 19.05 ATOM 3881CB VAL A 513 13.946 24.855 −45.428 1.00 19.18 ATOM 3882 CG1 VAL A 51313.041 24.064 −46.397 1.00 18.87 ATOM 3883 CG2 VAL A 513 15.042 23.938−44.895 1.00 20.56 ATOM 3884 C VAL A 513 13.536 26.979 −46.748 1.0017.98 ATOM 3885 O VAL A 513 12.910 27.768 −46.029 1.00 16.65 ATOM 3886 NLYS A 514 13.346 26.857 −48.063 1.00 17.91 ATOM 3887 CA LYS A 514 12.27927.583 −48.757 1.00 18.14 ATOM 3888 CB LYS A 514 12.845 28.712 −49.6381.00 17.66 ATOM 3889 CG LYS A 514 13.867 29.576 −48.945 1.00 19.26 ATOM3890 CD LYS A 514 14.197 30.839 −49.765 1.00 21.27 ATOM 3891 CE LYS A514 15.224 31.675 −49.001 1.00 26.06 ATOM 3892 NZ LYS A 514 15.46133.022 −49.626 1.00 28.70 ATOM 3893 C LYS A 514 11.494 26.621 −49.6251.00 18.22 ATOM 3894 O LYS A 514 11.949 25.502 −49.912 1.00 18.39 ATOM3895 N VAL A 515 10.304 27.037 −50.045 1.00 18.20 ATOM 3896 CA VAL A 5159.546 26.212 −50.980 1.00 19.00 ATOM 3897 CB VAL A 515 8.198 25.731−50.404 1.00 20.00 ATOM 3898 CG1 VAL A 515 7.403 26.904 −49.879 1.0021.01 ATOM 3899 CG2 VAL A 515 7.417 24.903 −51.447 1.00 20.10 ATOM 3900C VAL A 515 9.421 26.973 −52.302 1.00 18.89 ATOM 3901 O VAL A 515 9.07928.159 −52.317 1.00 18.67 ATOM 3902 N ALA A 516 9.781 26.295 −53.3901.00 19.87 ATOM 3903 CA ALA A 516 9.796 26.898 −54.732 1.00 20.38 ATOM3904 CB ALA A 516 11.177 26.768 −55.356 1.00 20.58 ATOM 3905 C ALA A 5168.789 26.110 −55.525 1.00 20.65 ATOM 3906 O ALA A 516 8.638 24.910−55.303 1.00 20.40 ATOM 3907 N GLY A 517 8.075 26.765 −56.430 1.00 20.80ATOM 3908 CA GLY A 517 7.092 26.039 −57.214 1.00 22.32 ATOM 3909 C GLY A517 6.536 26.853 −58.352 1.00 22.86 ATOM 3910 O GLY A 517 6.902 28.024−58.527 1.00 22.58 ATOM 3911 N ASN A 518 5.642 26.233 −59.116 1.00 24.66ATOM 3912 CA ASN A 518 5.201 26.817 −60.390 1.00 26.74 ATOM 3913 CB ASNA 518 4.670 25.754 −61.354 1.00 26.97 ATOM 3914 CG ASN A 518 3.38625.117 −60.872 1.00 31.69 ATOM 3915 OD1 ASN A 518 3.004 25.226 −59.6771.00 28.89 ATOM 3916 ND2 ASN A 518 2.707 24.419 −61.786 1.00 31.65 ATOM3917 C ASN A 518 4.199 27.937 −60.232 1.00 27.20 ATOM 3918 O ASN A 5184.154 28.822 −61.079 1.00 28.68 ATOM 3919 N ALA A 519 3.399 27.907−59.163 1.00 27.04 ATOM 3920 CA ALA A 519 2.424 28.978 −58.898 1.0026.47 ATOM 3921 CB ALA A 519 1.473 28.598 −57.747 1.00 27.02 ATOM 3922 CALA A 519 3.090 30.322 −58.629 1.00 26.35 ATOM 3923 O ALA A 519 4.22630.394 −58.135 1.00 25.27 ATOM 3924 N ALA A 520 2.369 31.394 −58.9541.00 26.25 ATOM 3925 CA ALA A 520 2.887 32.741 −58.784 1.00 26.77 ATOM3926 CB ALA A 520 1.872 33.775 −59.298 1.00 27.50 ATOM 3927 C ALA A 5203.250 33.004 −57.317 1.00 26.68 ATOM 3928 O ALA A 520 4.301 33.560−57.030 1.00 26.01 ATOM 3929 N ALA A 521 2.395 32.548 −56.399 1.00 26.75ATOM 3930 CA ALA A 521 2.628 32.712 −54.963 1.00 26.82 ATOM 3931 CB ALAA 521 1.395 32.251 −54.167 1.00 26.85 ATOM 3932 C ALA A 521 3.876 31.950−54.504 1.00 26.51 ATOM 3933 O ALA A 521 4.485 32.305 −53.494 1.00 26.63ATOM 3934 N LEU A 522 4.261 30.919 −55.259 1.00 26.79 ATOM 3935 CA LEU A522 5.452 30.113 −54.932 1.00 26.50 ATOM 3936 CB LEU A 522 5.185 28.626−55.155 1.00 26.64 ATOM 3937 CG LEU A 522 4.224 27.946 −54.169 1.0026.58 ATOM 3938 CD1 LEU A 522 4.049 26.489 −54.533 1.00 27.59 ATOM 3939CD2 LEU A 522 4.718 28.092 −52.730 1.00 28.08 ATOM 3940 C LEU A 5226.696 30.559 −55.709 1.00 26.54 ATOM 3941 O LEU A 522 7.779 29.987−55.547 1.00 25.56 ATOM 3942 N GLY A 523 6.518 31.575 −56.552 1.00 26.25ATOM 3943 CA GLY A 523 7.637 32.267 −57.199 1.00 26.16 ATOM 3944 C GLY A523 7.996 31.809 −58.607 1.00 26.84 ATOM 3945 O GLY A 523 9.029 32.227−59.152 1.00 25.81 ATOM 3946 N ASN A 524 7.162 30.946 −59.193 1.00 27.13ATOM 3947 CA ASN A 524 7.413 30.419 −60.539 1.00 27.74 ATOM 3948 CB ASNA 524 7.046 31.484 −61.591 1.00 28.43 ATOM 3949 CG ASN A 524 7.12330.960 −63.015 1.00 30.79 ATOM 3950 OD1 ASN A 524 6.856 29.780 −63.2851.00 30.80 ATOM 3951 ND2 ASN A 524 7.515 31.838 −63.936 1.00 33.61 ATOM3952 C ASN A 524 8.845 29.857 −60.710 1.00 28.44 ATOM 3953 O ASN A 5249.531 30.104 −61.720 1.00 27.82 ATOM 3954 N TRP A 525 9.280 29.111−59.693 1.00 27.99 ATOM 3955 CA TRP A 525 10.573 28.398 −59.659 1.0028.92 ATOM 3956 CB TRP A 525 10.787 27.507 −60.896 1.00 28.31 ATOM 3957CG TRP A 525 9.803 26.394 −61.060 1.00 27.68 ATOM 3958 CD1 TRP A 5258.902 26.247 −62.078 1.00 27.96 ATOM 3959 NE1 TRP A 525 8.166 25.106−61.907 1.00 27.55 ATOM 3960 CE2 TRP A 525 8.589 24.471 −60.762 1.0030.58 ATOM 3961 CD2 TRP A 525 9.609 25.277 −60.184 1.00 27.84 ATOM 3962CE3 TRP A 525 10.230 24.842 −59.001 1.00 26.50 ATOM 3963 CZ3 TRP A 5259.787 23.655 −58.411 1.00 27.55 ATOM 3964 CH2 TRP A 525 8.752 22.889−58.998 1.00 27.90 ATOM 3965 CZ2 TRP A 525 8.144 23.279 −60.168 1.0026.22 ATOM 3966 C TRP A 525 11.790 29.301 −59.452 1.00 29.66 ATOM 3967 OTRP A 525 12.921 28.804 −59.346 1.00 30.61 ATOM 3968 N SER A 526 11.57030.613 −59.380 1.00 30.33 ATOM 3969 CA SER A 526 12.645 31.536 −59.0041.00 31.13 ATOM 3970 CB SER A 526 12.213 32.993 −59.187 1.00 31.02 ATOM3971 OG SER A 526 13.166 33.838 −58.562 1.00 33.69 ATOM 3972 C SER A 52613.086 31.312 −57.560 1.00 31.29 ATOM 3973 O SER A 526 12.271 31.381−56.627 1.00 31.21 ATOM 3974 N THR A 527 14.373 31.049 −57.367 1.0031.34 ATOM 3975 CA THR A 527 14.880 30.794 −56.021 1.00 31.64 ATOM 3976CB THR A 527 16.259 30.098 −56.024 1.00 31.79 ATOM 3977 OG1 THR A 52717.217 30.931 −56.682 1.00 31.43 ATOM 3978 CG2 THR A 527 16.169 28.739−56.724 1.00 32.27 ATOM 3979 C THR A 527 14.911 32.045 −55.152 1.0031.99 ATOM 3980 O THR A 527 14.847 31.959 −53.922 1.00 32.36 ATOM 3981 NSER A 528 14.986 33.209 −55.787 1.00 31.79 ATOM 3982 CA SER A 528 14.92834.463 −55.054 1.00 32.02 ATOM 3983 CB SER A 528 15.517 35.615 −55.8851.00 32.70 ATOM 3984 OG SER A 528 14.712 35.882 −57.031 1.00 34.94 ATOM3985 C SER A 528 13.497 34.784 −54.579 1.00 31.23 ATOM 3986 O SER A 52813.330 35.435 −53.550 1.00 32.02 ATOM 3987 N ALA A 529 12.479 34.314−55.306 1.00 29.33 ATOM 3988 CA ALA A 529 11.093 34.506 −54.893 1.0027.60 ATOM 3989 CB ALA A 529 10.211 34.864 −56.086 1.00 27.34 ATOM 3990C ALA A 529 10.482 33.328 −54.112 1.00 26.40 ATOM 3991 O ALA A 529 9.31133.382 −53.754 1.00 26.54 ATOM 3992 N ALA A 530 11.268 32.286 −53.8421.00 25.12 ATOM 3993 CA ALA A 530 10.777 31.114 −53.096 1.00 24.19 ATOM3994 CB ALA A 530 11.855 30.063 −52.991 1.00 23.88 ATOM 3995 C ALA A 53010.336 31.555 −51.706 1.00 23.55 ATOM 3996 O ALA A 530 10.848 32.540−51.182 1.00 23.61 ATOM 3997 N VAL A 531 9.396 30.833 −51.110 1.00 22.18ATOM 3998 CA VAL A 531 8.851 31.248 −49.821 1.00 22.62 ATOM 3999 CB VALA 531 7.380 30.821 −49.677 1.00 22.81 ATOM 4000 CG1 VAL A 531 6.81531.335 −48.346 1.00 25.06 ATOM 4001 CG2 VAL A 531 6.551 31.353 −50.8861.00 25.38 ATOM 4002 C VAL A 531 9.659 30.646 −48.674 1.00 21.29 ATOM4003 O VAL A 531 9.768 29.425 −48.564 1.00 20.72 ATOM 4004 N ALA A 53210.215 31.493 −47.819 1.00 20.86 ATOM 4005 CA ALA A 532 11.008 30.999−46.698 1.00 20.26 ATOM 4006 CB ALA A 532 11.850 32.128 −46.084 1.0021.02 ATOM 4007 C ALA A 532 10.093 30.356 −45.646 1.00 20.51 ATOM 4008 OALA A 532 9.019 30.884 −45.337 1.00 20.05 ATOM 4009 N LEU A 533 10.51429.200 −45.129 1.00 19.00 ATOM 4010 CA LEU A 533 9.855 28.565 −43.9991.00 18.65 ATOM 4011 CB LEU A 533 9.901 27.029 −44.148 1.00 18.14 ATOM4012 CG LEU A 533 9.395 26.450 −45.483 1.00 19.25 ATOM 4013 CD1 LEU A533 9.385 24.923 −45.427 1.00 21.28 ATOM 4014 CD2 LEU A 533 8.030 26.980−45.894 1.00 18.58 ATOM 4015 C LEU A 533 10.541 29.014 −42.702 1.0018.95 ATOM 4016 O LEU A 533 11.622 29.648 −42.744 1.00 18.94 ATOM 4017 NASP A 534 9.905 28.715 −41.570 1.00 18.56 ATOM 4018 CA ASP A 534 10.38129.096 −40.238 1.00 18.36 ATOM 4019 CB ASP A 534 9.220 29.634 −39.3741.00 19.76 ATOM 4020 CG ASP A 534 8.757 30.992 −39.798 1.00 23.55 ATOM4021 OD1 ASP A 534 7.548 31.264 −39.659 1.00 26.14 ATOM 4022 OD2 ASP A534 9.600 31.774 −40.283 1.00 27.39 ATOM 4023 C ASP A 534 10.877 27.867−39.504 1.00 17.68 ATOM 4024 O ASP A 534 10.310 26.780 −39.667 1.0016.10 ATOM 4025 N ALA A 535 11.883 28.057 −38.654 1.00 17.05 ATOM 4026CA ALA A 535 12.405 26.950 −37.835 1.00 17.76 ATOM 4027 CB ALA A 53513.926 26.952 −37.832 1.00 17.96 ATOM 4028 C ALA A 535 11.872 27.027−36.403 1.00 17.82 ATOM 4029 O ALA A 535 12.482 26.490 −35.474 1.0018.36 ATOM 4030 N VAL A 536 10.745 27.706 −36.225 1.00 17.68 ATOM 4031CA VAL A 536 10.138 27.861 −34.898 1.00 18.65 ATOM 4032 CB VAL A 5368.824 28.719 −34.975 1.00 18.66 ATOM 4033 CG1 VAL A 536 7.805 28.123−35.971 1.00 19.98 ATOM 4034 CG2 VAL A 536 8.208 28.962 −33.570 1.0020.19 ATOM 4035 C VAL A 536 9.938 26.514 −34.155 1.00 18.51 ATOM 4036 OVAL A 536 10.124 26.437 −32.923 1.00 19.61 ATOM 4037 N ASN A 537 9.57025.468 −34.883 1.00 18.59 ATOM 4038 CA ASN A 537 9.344 24.154 −34.2611.00 19.07 ATOM 4039 CB ASN A 537 8.074 23.498 −34.816 1.00 19.28 ATOM4040 CG ASN A 537 6.800 24.252 −34.448 1.00 21.02 ATOM 4041 OD1 ASN A537 6.742 24.940 −33.435 1.00 24.12 ATOM 4042 ND2 ASN A 537 5.762 24.089−35.265 1.00 20.99 ATOM 4043 C ASN A 537 10.518 23.182 −34.445 1.0019.15 ATOM 4044 O ASN A 537 10.394 21.971 −34.196 1.00 18.60 ATOM 4045 NTYR A 538 11.653 23.699 −34.897 1.00 19.05 ATOM 4046 CA TYR A 538 12.76722.830 −35.234 1.00 20.32 ATOM 4047 CB TYR A 538 13.816 23.618 −36.0261.00 20.37 ATOM 4048 CG TYR A 538 14.916 22.747 −36.588 1.00 20.37 ATOM4049 CD1 TYR A 538 14.822 22.238 −37.886 1.00 20.39 ATOM 4050 CE1 TYR A538 15.853 21.436 −38.437 1.00 20.49 ATOM 4051 CZ TYR A 538 16.96121.137 −37.670 1.00 20.65 ATOM 4052 OH TYR A 538 17.946 20.341 −38.2181.00 21.53 ATOM 4053 CE2 TYR A 538 17.066 21.602 −36.361 1.00 22.05 ATOM4054 CD2 TYR A 538 16.043 22.418 −35.825 1.00 21.95 ATOM 4055 C TYR A538 13.436 22.209 −33.981 1.00 21.29 ATOM 4056 O TYR A 538 13.733 22.919−33.036 1.00 21.89 ATOM 4057 N ALA A 539 13.695 20.902 −34.014 1.0022.15 ATOM 4058 CA ALA A 539 14.646 20.258 −33.083 1.00 23.67 ATOM 4059CB ALA A 539 13.909 19.536 −31.976 1.00 23.94 ATOM 4060 C ALA A 53915.545 19.289 −33.849 1.00 24.35 ATOM 4061 O ALA A 539 15.117 18.698−34.833 1.00 23.63 ATOM 4062 N ASP A 540 16.793 19.118 −33.405 1.0025.69 ATOM 4063 CA ASP A 540 17.722 18.196 −34.099 1.00 27.42 ATOM 4064CB ASP A 540 19.044 18.051 −33.339 1.00 28.62 ATOM 4065 CG ASP A 54019.724 19.368 −33.140 1.00 33.80 ATOM 4066 OD1 ASP A 540 19.875 20.115−34.147 1.00 36.83 ATOM 4067 OD2 ASP A 540 20.080 19.663 −31.970 1.0040.32 ATOM 4068 C ASP A 540 17.150 16.818 −34.400 1.00 26.63 ATOM 4069 OASP A 540 17.386 16.277 −35.485 1.00 27.28 ATOM 4070 N ASN A 541 16.40316.247 −33.458 1.00 25.08 ATOM 4071 CA ASN A 541 15.827 14.922 −33.6871.00 24.03 ATOM 4072 CB ASN A 541 15.988 14.032 −32.441 1.00 25.14 ATOM4073 CG ASN A 541 15.337 14.623 −31.191 1.00 27.81 ATOM 4074 OD1 ASN A541 15.366 14.001 −30.118 1.00 31.57 ATOM 4075 ND2 ASN A 541 14.77115.824 −31.306 1.00 28.81 ATOM 4076 C ASN A 541 14.349 14.991 −34.1181.00 22.46 ATOM 4077 O ASN A 541 13.660 13.979 −34.172 1.00 22.35 ATOM4078 N HIS A 542 13.871 16.197 −34.403 1.00 19.85 ATOM 4079 CA HIS A 54212.580 16.354 −35.083 1.00 18.22 ATOM 4080 CB HIS A 542 11.426 16.392−34.062 1.00 16.89 ATOM 4081 CG HIS A 542 10.071 16.341 −34.699 1.0017.32 ATOM 4082 ND1 HIS A 542 9.211 17.417 −34.711 1.00 16.28 ATOM 4083CE1 HIS A 542 8.111 17.088 −35.371 1.00 14.64 ATOM 4084 NE2 HIS A 5428.217 15.834 −35.765 1.00 15.41 ATOM 4085 CD2 HIS A 542 9.435 15.340−35.358 1.00 15.14 ATOM 4086 C HIS A 542 12.662 17.650 −35.902 1.0017.48 ATOM 4087 O HIS A 542 12.198 18.698 −35.446 1.00 17.79 ATOM 4088 NPRO A 543 13.324 17.584 −37.083 1.00 17.00 ATOM 4089 CA PRO A 543 13.79718.752 −37.832 1.00 17.09 ATOM 4090 CB PRO A 543 14.948 18.164 −38.6771.00 16.57 ATOM 4091 CG PRO A 543 14.472 16.759 −38.994 1.00 18.22 ATOM4092 CD PRO A 543 13.676 16.321 −37.764 1.00 17.39 ATOM 4093 C PRO A 54312.718 19.435 −38.691 1.00 16.46 ATOM 4094 O PRO A 543 12.811 19.497−39.922 1.00 17.09 ATOM 4095 N LEU A 544 11.726 19.987 −38.009 1.0015.36 ATOM 4096 CA LEU A 544 10.534 20.535 −38.636 1.00 15.10 ATOM 4097CB LEU A 544 9.341 20.376 −37.672 1.00 15.29 ATOM 4098 CG LEU A 5447.968 20.927 −38.134 1.00 16.53 ATOM 4099 CD1 LEU A 544 7.524 20.364−39.494 1.00 15.92 ATOM 4100 CD2 LEU A 544 6.900 20.700 −37.062 1.0014.59 ATOM 4101 C LEU A 544 10.694 22.018 −39.025 1.00 15.09 ATOM 4102 OLEU A 544 11.037 22.851 −38.197 1.00 15.34 ATOM 4103 N TRP A 545 10.45622.298 −40.303 1.00 15.03 ATOM 4104 CA TRP A 545 10.327 23.637 −40.8431.00 15.36 ATOM 4105 CB TRP A 545 11.288 23.790 −42.023 1.00 15.40 ATOM4106 CG TRP A 545 12.758 23.921 −41.663 1.00 15.62 ATOM 4107 CD1 TRP A545 13.653 22.903 −41.384 1.00 16.85 ATOM 4108 NE1 TRP A 545 14.90623.437 −41.129 1.00 18.69 ATOM 4109 CE2 TRP A 545 14.837 24.803 −41.2461.00 17.78 ATOM 4110 CD2 TRP A 545 13.498 25.140 −41.584 1.00 17.01 ATOM4111 CE3 TRP A 545 13.163 26.488 −41.777 1.00 17.17 ATOM 4112 CZ3 TRP A545 14.165 27.456 −41.637 1.00 18.29 ATOM 4113 CH2 TRP A 545 15.48327.085 −41.295 1.00 17.22 ATOM 4114 CZ2 TRP A 545 15.835 25.767 −41.1111.00 19.09 ATOM 4115 C TRP A 545 8.907 23.832 −41.359 1.00 15.14 ATOM4116 O TRP A 545 8.327 22.933 −41.986 1.00 14.45 ATOM 4117 N ILE A 5468.362 25.025 −41.149 1.00 15.19 ATOM 4118 CA ILE A 546 6.938 25.244−41.428 1.00 16.51 ATOM 4119 CB ILE A 546 6.107 24.988 −40.130 1.0017.39 ATOM 4120 CG1 ILE A 546 4.615 24.852 −40.420 1.00 20.44 ATOM 4121CD1 ILE A 546 3.882 23.992 −39.392 1.00 23.59 ATOM 4122 CG2 ILE A 5466.391 26.064 −39.050 1.00 17.70 ATOM 4123 C ILE A 546 6.674 26.635−42.006 1.00 16.75 ATOM 4124 O ILE A 546 7.352 27.593 −41.647 1.00 15.81ATOM 4125 N ALA A 547 5.716 26.743 −42.925 1.00 16.88 ATOM 4126 CA ALA A547 5.197 28.057 −43.279 1.00 17.94 ATOM 4127 CB ALA A 547 6.222 28.893−43.931 1.00 21.49 ATOM 4128 C ALA A 547 4.009 27.919 −44.167 1.00 18.29ATOM 4129 O ALA A 547 3.727 26.828 −44.655 1.00 18.52 ATOM 4130 N THR A548 3.316 29.031 −44.362 1.00 17.99 ATOM 4131 CA THR A 548 1.970 29.017−44.919 1.00 18.72 ATOM 4132 CB THR A 548 0.929 29.419 −43.855 1.0018.65 ATOM 4133 OG1 THR A 548 1.000 28.500 −42.751 1.00 19.46 ATOM 4134CG2 THR A 548 −0.491 29.379 −44.438 1.00 18.79 ATOM 4135 C THR A 5481.865 29.960 −46.104 1.00 19.60 ATOM 4136 O THR A 548 2.347 31.090−46.040 1.00 19.83 ATOM 4137 N VAL A 549 1.227 29.485 −47.164 1.00 20.41ATOM 4138 CA VAL A 549 1.048 30.280 −48.389 1.00 22.36 ATOM 4139 CB VALA 549 1.944 29.722 −49.537 1.00 22.77 ATOM 4140 CG1 VAL A 549 1.71730.491 −50.845 1.00 25.80 ATOM 4141 CG2 VAL A 549 3.429 29.781 −49.1481.00 24.78 ATOM 4142 C VAL A 549 −0.399 30.119 −48.800 1.00 22.21 ATOM4143 O VAL A 549 −0.943 29.018 −48.719 1.00 21.56 ATOM 4144 N ASN A 550−1.028 31.211 −49.240 1.00 22.80 ATOM 4145 CA ASN A 550 −2.356 31.107−49.831 1.00 23.95 ATOM 4146 CB ASN A 550 −3.114 32.411 −49.649 1.0024.39 ATOM 4147 CG ASN A 550 −3.367 32.706 −48.201 1.00 27.42 ATOM 4148OD1 ASN A 550 −3.811 31.838 −47.462 1.00 28.69 ATOM 4149 ND2 ASN A 550−3.041 33.911 −47.771 1.00 31.89 ATOM 4150 C ASN A 550 −2.278 30.733−51.294 1.00 23.98 ATOM 4151 O ASN A 550 −1.598 31.400 −52.065 1.0024.73 ATOM 4152 N LEU A 551 −2.973 29.667 −51.662 1.00 24.24 ATOM 4153CA LEU A 551 −3.016 29.180 −53.020 1.00 24.78 ATOM 4154 CB LEU A 551−2.348 27.797 −53.135 1.00 25.00 ATOM 4155 CG LEU A 551 −0.858 27.721−52.787 1.00 25.19 ATOM 4156 CD1 LEU A 551 −0.356 26.284 −52.803 1.0027.45 ATOM 4157 CD2 LEU A 551 −0.018 28.613 −53.718 1.00 26.73 ATOM 4158C LEU A 551 −4.471 29.104 −53.488 1.00 25.46 ATOM 4159 O LEU A 551−5.393 29.018 −52.675 1.00 24.27 ATOM 4160 N GLU A 552 −4.661 29.148−54.804 1.00 26.15 ATOM 4161 CA GLU A 552 −6.004 29.165 −55.366 1.0028.20 ATOM 4162 CB GLU A 552 −5.955 29.639 −56.823 1.00 28.30 ATOM 4163CG GLU A 552 −7.326 29.739 −57.494 1.00 32.28 ATOM 4164 CD GLU A 552−7.250 30.281 −58.926 1.00 33.44 ATOM 4165 OE1 GLU A 552 −8.110 31.126−59.274 1.00 41.80 ATOM 4166 OE2 GLU A 552 −6.340 29.873 −59.695 1.0039.14 ATOM 4167 C GLU A 552 −6.610 27.768 −55.253 1.00 26.97 ATOM 4168 OGLU A 552 −5.979 26.783 −55.622 1.00 26.49 ATOM 4169 N ALA A 553 −7.82227.684 −54.723 1.00 27.21 ATOM 4170 CA ALA A 553 −8.502 26.399 −54.6031.00 27.89 ATOM 4171 CB ALA A 553 −9.876 26.574 −53.953 1.00 28.37 ATOM4172 C ALA A 553 −8.637 25.773 −55.979 1.00 28.57 ATOM 4173 O ALA A 553−8.900 26.477 −56.952 1.00 28.83 ATOM 4174 N GLY A 554 −8.438 24.465−56.064 1.00 28.44 ATOM 4175 CA GLY A 554 −8.556 23.753 −57.330 1.0029.66 ATOM 4176 C GLY A 554 −7.274 23.693 −58.145 1.00 30.04 ATOM 4177 OGLY A 554 −7.122 22.814 −59.000 1.00 30.33 ATOM 4178 N ASP A 555 −6.34724.606 −57.869 1.00 30.23 ATOM 4179 CA ASP A 555 −5.098 24.716 −58.6301.00 30.61 ATOM 4180 CB ASP A 555 −4.313 25.939 −58.161 1.00 30.85 ATOM4181 CG ASP A 555 −3.382 26.503 −59.236 1.00 34.49 ATOM 4182 OD1 ASP A555 −3.441 26.053 −60.407 1.00 37.19 ATOM 4183 OD2 ASP A 555 −2.57227.408 −58.901 1.00 36.59 ATOM 4184 C ASP A 555 −4.238 23.467 −58.4861.00 30.14 ATOM 4185 O ASP A 555 −4.156 22.882 −57.419 1.00 30.07 ATOM4186 N VAL A 556 −3.602 23.046 −59.572 1.00 29.55 ATOM 4187 CA VAL A 556−2.628 21.963 −59.492 1.00 28.74 ATOM 4188 CB VAL A 556 −2.732 20.987−60.680 1.00 29.35 ATOM 4189 CG1 VAL A 556 −1.666 19.877 −60.569 1.0028.99 ATOM 4190 CG2 VAL A 556 −4.125 20.365 −60.720 1.00 30.44 ATOM 4191C VAL A 556 −1.261 22.623 −59.448 1.00 28.04 ATOM 4192 O VAL A 556−0.869 23.336 −60.384 1.00 27.25 ATOM 4193 N VAL A 557 −0.544 22.389−58.352 1.00 26.81 ATOM 4194 CA VAL A 557 0.739 23.040 −58.100 1.0026.14 ATOM 4195 CB VAL A 557 0.690 23.859 −56.759 1.00 26.71 ATOM 4196CG1 VAL A 557 2.073 24.140 −56.219 1.00 27.82 ATOM 4197 CG2 VAL A 557−0.088 25.175 −56.952 1.00 27.34 ATOM 4198 C VAL A 557 1.856 21.999−58.092 1.00 25.64 ATOM 4199 O VAL A 557 1.646 20.845 −57.693 1.00 25.24ATOM 4200 N GLU A 558 3.035 22.409 −58.553 1.00 24.16 ATOM 4201 CA GLU A558 4.223 21.579 −58.516 1.00 24.38 ATOM 4202 CB GLU A 558 4.737 21.296−59.933 1.00 24.86 ATOM 4203 CG GLU A 558 4.064 20.108 −60.606 1.0026.34 ATOM 4204 CD GLU A 558 4.670 19.790 −61.962 1.00 27.68 ATOM 4205OE1 GLU A 558 5.917 19.684 −62.065 1.00 30.56 ATOM 4206 OE2 GLU A 5583.883 19.638 −62.915 1.00 32.45 ATOM 4207 C GLU A 558 5.262 22.337−57.730 1.00 22.54 ATOM 4208 O GLU A 558 5.389 23.550 −57.883 1.00 22.93ATOM 4209 N TYR A 559 5.992 21.640 −56.867 1.00 21.45 ATOM 4210 CA TYR A559 6.927 22.346 −55.995 1.00 19.96 ATOM 4211 CB TYR A 559 6.188 22.972−54.784 1.00 19.42 ATOM 4212 CG TYR A 559 5.624 21.952 −53.796 1.0018.65 ATOM 4213 CD1 TYR A 559 6.383 21.524 −52.703 1.00 18.64 ATOM 4214CE1 TYR A 559 5.887 20.595 −51.794 1.00 19.09 ATOM 4215 CZ TYR A 5594.614 20.090 −51.955 1.00 18.58 ATOM 4216 OH TYR A 559 4.135 19.160−51.056 1.00 20.55 ATOM 4217 CE2 TYR A 559 3.819 20.493 −53.024 1.0018.22 ATOM 4218 CD2 TYR A 559 4.335 21.438 −53.946 1.00 18.82 ATOM 4219C TYR A 559 8.066 21.445 −55.541 1.00 20.13 ATOM 4220 O TYR A 559 8.00820.215 −55.679 1.00 20.26 ATOM 4221 N LYS A 560 9.098 22.079 −54.9951.00 19.51 ATOM 4222 CA LYS A 560 10.208 21.379 −54.349 1.00 19.72 ATOM4223 CB LYS A 560 11.410 21.175 −55.282 1.00 19.66 ATOM 4224 CG LYS A560 11.390 19.870 −56.058 1.00 22.20 ATOM 4225 CD LYS A 560 12.76719.633 −56.714 1.00 24.46 ATOM 4226 CE LYS A 560 12.781 18.341 −57.5311.00 26.96 ATOM 4227 NZ LYS A 560 14.189 18.050 −57.980 1.00 26.97 ATOM4228 C LYS A 560 10.680 22.257 −53.234 1.00 18.68 ATOM 4229 O LYS A 56010.583 23.484 −53.318 1.00 19.68 ATOM 4230 N TYR A 561 11.240 21.640−52.206 1.00 17.87 ATOM 4231 CA TYR A 561 11.927 22.420 −51.187 1.0017.82 ATOM 4232 CB TYR A 561 11.921 21.690 −49.840 1.00 17.42 ATOM 4233CG TYR A 561 10.518 21.449 −49.346 1.00 16.12 ATOM 4234 CD1 TYR A 5619.831 20.276 −49.661 1.00 15.94 ATOM 4235 CE1 TYR A 561 8.511 20.062−49.199 1.00 15.79 ATOM 4236 CZ TYR A 561 7.897 21.050 −48.456 1.0015.89 ATOM 4237 OH TYR A 561 6.614 20.889 −47.981 1.00 17.17 ATOM 4238CE2 TYR A 561 8.557 22.224 −48.164 1.00 16.19 ATOM 4239 CD2 TYR A 5619.856 22.430 −48.625 1.00 17.57 ATOM 4240 C TYR A 561 13.360 22.650−51.607 1.00 18.63 ATOM 4241 O TYR A 561 13.963 21.786 −52.265 1.0018.40 ATOM 4242 N ILE A 562 13.904 23.792 −51.201 1.00 18.56 ATOM 4243CA ILE A 562 15.322 24.085 −51.434 1.00 20.18 ATOM 4244 CB ILE A 56215.524 25.247 −52.419 1.00 19.65 ATOM 4245 CG1 ILE A 562 14.837 26.520−51.896 1.00 21.64 ATOM 4246 CD1 ILE A 562 15.074 27.789 −52.741 1.0021.24 ATOM 4247 CG2 ILE A 562 15.017 24.829 −53.797 1.00 20.44 ATOM 4248C ILE A 562 15.971 24.446 −50.128 1.00 20.56 ATOM 4249 O ILE A 56215.316 24.956 −49.229 1.00 19.84 ATOM 4250 N ASN A 563 17.254 24.134−50.029 1.00 21.41 ATOM 4251 CA ASN A 563 18.076 24.467 −48.886 1.0022.99 ATOM 4252 CB ASN A 563 18.833 23.209 −48.435 1.00 22.69 ATOM 4253CG ASN A 563 19.629 23.433 −47.156 1.00 25.48 ATOM 4254 OD1 ASN A 56320.203 24.492 −46.965 1.00 28.81 ATOM 4255 ND2 ASN A 563 19.669 22.436−46.285 1.00 25.82 ATOM 4256 C ASN A 563 19.039 25.561 −49.372 1.0024.66 ATOM 4257 O ASN A 563 19.794 25.326 −50.323 1.00 24.16 ATOM 4258 NVAL A 564 18.977 26.749 −48.780 1.00 26.20 ATOM 4259 CA VAL A 564 19.87727.837 −49.206 1.00 28.74 ATOM 4260 CB VAL A 564 19.156 29.091 −49.8321.00 28.91 ATOM 4261 CG1 VAL A 564 19.461 30.408 −49.087 1.00 31.10 ATOM4262 CG2 VAL A 564 17.655 28.836 −50.079 1.00 28.92 ATOM 4263 C VAL A564 20.886 28.181 −48.122 1.00 29.56 ATOM 4264 O VAL A 564 20.538 28.320−46.954 1.00 29.10 ATOM 4265 N GLY A 565 22.150 28.266 −48.527 1.0032.02 ATOM 4266 CA GLY A 565 23.252 28.423 −47.577 1.00 34.92 ATOM 4267C GLY A 565 23.539 29.876 −47.248 1.00 37.47 ATOM 4268 O GLY A 56522.969 30.788 −47.871 1.00 37.66 ATOM 4269 N GLN A 566 24.419 30.098−46.267 1.00 39.85 ATOM 4270 CA GLN A 566 24.897 31.456 −45.926 1.0042.75 ATOM 4271 CB GLN A 566 26.054 31.398 −44.918 1.00 42.83 ATOM 4272CG GLN A 566 25.727 30.761 −43.565 1.00 44.99 ATOM 4273 CD GLN A 56626.940 30.689 −42.626 1.00 44.88 ATOM 4274 OE1 GLN A 566 27.972 30.089−42.958 1.00 47.67 ATOM 4275 NE2 GLN A 566 26.810 31.293 −41.441 1.0047.62 ATOM 4276 C GLN A 566 25.373 32.195 −47.181 1.00 43.30 ATOM 4277 OGLN A 566 25.052 33.365 −47.389 1.00 44.01 ATOM 4278 N ASP A 567 26.11831.479 −48.023 1.00 44.30 ATOM 4279 CA ASP A 567 26.739 32.029 −49.2361.00 44.62 ATOM 4280 CB ASP A 567 27.916 31.139 −49.650 1.00 45.13 ATOM4281 CG ASP A 567 27.492 29.702 −49.966 1.00 47.62 ATOM 4282 OD1 ASP A567 26.421 29.255 −49.485 1.00 48.73 ATOM 4283 OD2 ASP A 567 28.24529.010 −50.693 1.00 50.47 ATOM 4284 C ASP A 567 25.776 32.197 −50.4211.00 44.04 ATOM 4285 O ASP A 567 26.196 32.575 −51.522 1.00 44.36 ATOM4286 N GLY A 568 24.497 31.899 −50.205 1.00 42.85 ATOM 4287 CA GLY A 56823.488 32.045 −51.247 1.00 41.38 ATOM 4288 C GLY A 568 23.359 30.851−52.177 1.00 40.41 ATOM 4289 O GLY A 568 22.496 30.854 −53.054 1.0040.62 ATOM 4290 N SER A 569 24.195 29.827 −51.990 1.00 39.02 ATOM 4291CA SER A 569 24.137 28.623 −52.827 1.00 37.72 ATOM 4292 CB SER A 56925.365 27.746 −52.600 1.00 38.07 ATOM 4293 OG SER A 569 25.454 27.359−51.238 1.00 39.18 ATOM 4294 C SER A 569 22.868 27.819 −52.540 1.0036.72 ATOM 4295 O SER A 569 22.474 27.672 −51.382 1.00 36.47 ATOM 4296 NVAL A 570 22.222 27.313 −53.583 1.00 35.25 ATOM 4297 CA VAL A 570 20.98826.558 −53.365 1.00 34.26 ATOM 4298 CB VAL A 570 19.709 27.243 −53.9871.00 34.42 ATOM 4299 CG1 VAL A 570 18.992 26.358 −55.020 1.00 35.11 ATOM4300 CG2 VAL A 570 20.010 28.651 −54.506 1.00 35.12 ATOM 4301 C VAL A570 21.113 25.080 −53.718 1.00 33.29 ATOM 4302 O VAL A 570 21.735 24.694−54.714 1.00 32.88 ATOM 4303 N THR A 571 20.515 24.261 −52.864 1.0031.66 ATOM 4304 CA THR A 571 20.480 22.825 −53.021 1.00 30.87 ATOM 4305CB THR A 571 21.016 22.146 −51.752 1.00 30.96 ATOM 4306 OG1 THR A 57122.311 22.686 −51.442 1.00 33.18 ATOM 4307 CG2 THR A 571 21.117 20.637−51.935 1.00 30.95 ATOM 4308 C THR A 571 19.018 22.473 −53.210 1.0029.72 ATOM 4309 O THR A 571 18.186 22.832 −52.373 1.00 28.84 ATOM 4310 NTRP A 572 18.697 21.846 −54.337 1.00 29.01 ATOM 4311 CA TRP A 572 17.33121.388 −54.589 1.00 28.49 ATOM 4312 CB TRP A 572 17.004 21.367 −56.0861.00 28.99 ATOM 4313 CG TRP A 572 16.950 22.690 −56.739 1.00 29.65 ATOM4314 CD1 TRP A 572 18.014 23.419 −57.217 1.00 30.67 ATOM 4315 NE1 TRP A572 17.564 24.603 −57.769 1.00 31.84 ATOM 4316 CE2 TRP A 572 16.19624.655 −57.668 1.00 30.45 ATOM 4317 CD2 TRP A 572 15.770 23.464 −57.0281.00 30.30 ATOM 4318 CE3 TRP A 572 14.398 23.274 −56.791 1.00 29.43 ATOM4319 CZ3 TRP A 572 13.502 24.266 −57.205 1.00 30.24 ATOM 4320 CH2 TRP A572 13.959 25.437 −57.846 1.00 30.31 ATOM 4321 CZ2 TRP A 572 15.29825.649 −58.083 1.00 29.71 ATOM 4322 C TRP A 572 17.205 19.991 −54.0311.00 28.21 ATOM 4323 O TRP A 572 18.168 19.212 −54.060 1.00 27.50 ATOM4324 N GLU A 573 16.033 19.647 −53.499 1.00 27.69 ATOM 4325 CA GLU A 57315.819 18.251 −53.123 1.00 27.17 ATOM 4326 CB GLU A 573 14.586 18.074−52.222 1.00 27.18 ATOM 4327 CG GLU A 573 13.287 18.406 −52.901 1.0026.04 ATOM 4328 CD GLU A 573 12.059 18.111 −52.028 1.00 26.20 ATOM 4329OE1 GLU A 573 12.112 17.224 −51.141 1.00 25.84 ATOM 4330 OE2 GLU A 57311.032 18.764 −52.264 1.00 22.24 ATOM 4331 C GLU A 573 15.725 17.419−54.405 1.00 27.91 ATOM 4332 O GLU A 573 15.498 17.957 −55.497 1.0026.94 ATOM 4333 N SER A 574 15.907 16.108 −54.267 1.00 28.48 ATOM 4334CA SER A 574 15.880 15.201 −55.410 1.00 29.59 ATOM 4335 CB SER A 57416.296 13.805 −54.975 1.00 29.73 ATOM 4336 OG SER A 574 17.609 13.875−54.449 1.00 32.74 ATOM 4337 C SER A 574 14.526 15.134 −56.095 1.0029.70 ATOM 4338 O SER A 574 13.500 15.482 −55.513 1.00 29.12 ATOM 4339 NASP A 575 14.544 14.669 −57.339 1.00 29.54 ATOM 4340 CA ASP A 575 13.33714.435 −58.109 1.00 29.98 ATOM 4341 CB ASP A 575 13.705 14.000 −59.5341.00 30.69 ATOM 4342 CG ASP A 575 14.324 15.125 −60.331 1.00 33.96 ATOM4343 OD1 ASP A 575 14.056 16.299 −59.997 1.00 36.25 ATOM 4344 OD2 ASP A575 15.083 14.846 −61.290 1.00 37.94 ATOM 4345 C ASP A 575 12.519 13.358−57.428 1.00 28.93 ATOM 4346 O ASP A 575 13.050 12.633 −56.600 1.0028.34 ATOM 4347 N PRO A 576 11.217 13.267 −57.760 1.00 28.58 ATOM 4348CA PRO A 576 10.469 14.173 −58.650 1.00 27.92 ATOM 4349 CB PRO A 5769.319 13.294 −59.131 1.00 28.52 ATOM 4350 CG PRO A 576 9.053 12.378−57.954 1.00 28.29 ATOM 4351 CD PRO A 576 10.377 12.159 −57.267 1.0028.76 ATOM 4352 C PRO A 576 9.894 15.397 −57.938 1.00 27.25 ATOM 4353 OPRO A 576 9.887 15.452 −56.703 1.00 28.13 ATOM 4354 N ASN A 577 9.39416.360 −58.707 1.00 25.62 ATOM 4355 CA ASN A 577 8.612 17.449 −58.1291.00 24.83 ATOM 4356 CB ASN A 577 8.013 18.336 −59.224 1.00 24.90 ATOM4357 CG ASN A 577 9.055 19.184 −59.913 1.00 25.61 ATOM 4358 OD1 ASN A577 10.176 19.321 −59.423 1.00 25.83 ATOM 4359 ND2 ASN A 577 8.69319.756 −61.060 1.00 25.07 ATOM 4360 C ASN A 577 7.466 16.868 −57.3221.00 24.22 ATOM 4361 O ASN A 577 6.949 15.798 −57.672 1.00 23.69 ATOM4362 N HIS A 578 7.057 17.562 −56.259 1.00 23.20 ATOM 4363 CA HIS A 5785.830 17.179 −55.570 1.00 22.96 ATOM 4364 CB HIS A 578 5.734 17.844−54.200 1.00 22.09 ATOM 4365 CG HIS A 578 6.874 17.538 −53.285 1.0021.80 ATOM 4366 ND1 HIS A 578 6.809 16.558 −52.318 1.00 21.62 ATOM 4367CE1 HIS A 578 7.948 16.530 −51.645 1.00 21.36 ATOM 4368 NE2 HIS A 5788.743 17.465 −52.133 1.00 20.16 ATOM 4369 CD2 HIS A 578 8.096 18.109−53.160 1.00 19.78 ATOM 4370 C HIS A 578 4.697 17.707 −56.429 1.00 23.58ATOM 4371 O HIS A 578 4.814 18.794 −56.976 1.00 23.64 ATOM 4372 N THR A579 3.603 16.955 −56.534 1.00 23.98 ATOM 4373 CA THR A 579 2.426 17.448−57.254 1.00 25.34 ATOM 4374 CB THR A 579 2.092 16.568 −58.477 1.0026.08 ATOM 4375 OG1 THR A 579 3.162 16.672 −59.429 1.00 29.14 ATOM 4376CG2 THR A 579 0.749 16.979 −59.126 1.00 26.14 ATOM 4377 C THR A 5791.259 17.480 −56.291 1.00 25.05 ATOM 4378 O THR A 579 0.977 16.487−55.629 1.00 25.73 ATOM 4379 N TYR A 580 0.591 18.619 −56.213 1.00 25.23ATOM 4380 CA TYR A 580 −0.450 18.802 −55.211 1.00 25.68 ATOM 4381 CB TYRA 580 0.098 19.556 −53.976 1.00 25.88 ATOM 4382 CG TYR A 580 −0.93119.763 −52.866 1.00 26.21 ATOM 4383 CD1 TYR A 580 −1.284 21.048 −52.4291.00 26.22 ATOM 4384 CE1 TYR A 580 −2.256 21.233 −51.399 1.00 27.53 ATOM4385 CZ TYR A 580 −2.860 20.111 −50.841 1.00 27.08 ATOM 4386 OH TYR A580 −3.806 20.207 −49.844 1.00 27.88 ATOM 4387 CE2 TYR A 580 −2.51018.841 −51.264 1.00 26.97 ATOM 4388 CD2 TYR A 580 −1.562 18.671 −52.2761.00 26.29 ATOM 4389 C TYR A 580 −1.634 19.523 −55.828 1.00 25.65 ATOM4390 O TYR A 580 −1.490 20.596 −56.403 1.00 25.29 ATOM 4391 N THR A 581−2.813 18.915 −55.732 1.00 25.95 ATOM 4392 CA THR A 581 −4.015 19.629−56.117 1.00 25.78 ATOM 4393 CB THR A 581 −5.016 18.700 −56.806 1.0026.75 ATOM 4394 OG1 THR A 581 −4.332 18.000 −57.855 1.00 26.57 ATOM 4395CG2 THR A 581 −6.189 19.498 −57.397 1.00 27.62 ATOM 4396 C THR A 581−4.627 20.285 −54.874 1.00 25.36 ATOM 4397 O THR A 581 −5.024 19.595−53.935 1.00 25.22 ATOM 4398 N VAL A 582 −4.685 21.615 −54.880 1.0024.54 ATOM 4399 CA VAL A 582 −5.255 22.382 −53.777 1.00 24.15 ATOM 4400CB VAL A 582 −5.006 23.915 −53.953 1.00 24.21 ATOM 4401 CG1 VAL A 582−5.472 24.700 −52.744 1.00 23.87 ATOM 4402 CG2 VAL A 582 −3.514 24.218−54.219 1.00 25.33 ATOM 4403 C VAL A 582 −6.759 22.063 −53.706 1.0024.32 ATOM 4404 O VAL A 582 −7.478 22.204 −54.700 1.00 23.26 ATOM 4405 NPRO A 583 −7.236 21.587 −52.546 1.00 24.10 ATOM 4406 CA PRO A 583 −8.66521.230 −52.476 1.00 24.34 ATOM 4407 CB PRO A 583 −8.865 20.763 −51.0221.00 24.33 ATOM 4408 CG PRO A 583 −7.516 20.538 −50.468 1.00 25.16 ATOM4409 CD PRO A 583 −6.508 21.310 −51.294 1.00 24.72 ATOM 4410 C PRO A 583−9.597 22.404 −52.768 1.00 24.50 ATOM 4411 O PRO A 583 −9.262 23.558−52.487 1.00 23.93 ATOM 4412 N ALA A 584 −10.756 22.104 −53.350 1.0024.61 ATOM 4413 CA ALA A 584 −11.817 23.084 −53.477 1.00 24.77 ATOM 4414CB ALA A 584 −12.065 23.439 −54.943 1.00 25.29 ATOM 4415 C ALA A 584−13.036 22.434 −52.847 1.00 24.94 ATOM 4416 O ALA A 584 −13.922 21.932−53.537 1.00 25.03 ATOM 4417 N VAL A 585 −13.052 22.406 −51.517 1.0024.24 ATOM 4418 CA VAL A 585 −14.075 21.673 −50.776 1.00 23.75 ATOM 4419CB VAL A 585 −13.465 20.452 −50.029 1.00 24.50 ATOM 4420 CG1 VAL A 585−14.515 19.770 −49.151 1.00 24.48 ATOM 4421 CG2 VAL A 585 −12.863 19.447−51.026 1.00 25.32 ATOM 4422 C VAL A 585 −14.707 22.639 −49.781 1.0023.20 ATOM 4423 O VAL A 585 −13.999 23.347 −49.065 1.00 22.13 ATOM 4424N ALA A 586 −16.044 22.679 −49.739 1.00 22.43 ATOM 4425 CA ALA A 586−16.749 23.546 −48.804 1.00 21.79 ATOM 4426 CB ALA A 586 −18.240 23.212−48.820 1.00 22.19 ATOM 4427 C ALA A 586 −16.160 23.324 −47.389 1.0021.57 ATOM 4428 O ALA A 586 −15.954 22.180 −46.990 1.00 20.89 ATOM 4429N CYS A 587 −15.872 24.414 −46.679 1.00 21.59 ATOM 4430 CA CYS A 587−15.388 24.379 −45.268 1.00 21.68 ATOM 4431 CB CYS A 587 −16.131 23.323−44.441 1.00 22.08 ATOM 4432 SG CYS A 587 −17.952 23.374 −44.507 1.0023.60 ATOM 4433 C CYS A 587 −13.886 24.129 −45.094 1.00 21.27 ATOM 4434O CYS A 587 −13.386 24.225 −43.980 1.00 21.08 ATOM 4435 N VAL A 588−13.178 23.780 −46.170 1.00 20.53 ATOM 4436 CA VAL A 588 −11.742 23.499−46.085 1.00 20.52 ATOM 4437 CB VAL A 588 −11.351 22.268 −46.958 1.0020.47 ATOM 4438 CG1 VAL A 588 −9.846 21.959 −46.844 1.00 20.87 ATOM 4439CG2 VAL A 588 −12.163 21.042 −46.549 1.00 20.51 ATOM 4440 C VAL A 588−10.949 24.731 −46.504 1.00 20.59 ATOM 4441 O VAL A 588 −10.699 24.950−47.705 1.00 21.88 ATOM 4442 N THR A 589 −10.533 25.522 −45.528 1.0019.56 ATOM 4443 CA THR A 589 −9.903 26.807 −45.795 1.00 19.48 ATOM 4444CB THR A 589 −10.595 27.914 −44.988 1.00 20.13 ATOM 4445 OG1 THR A 589−10.527 27.565 −43.592 1.00 21.49 ATOM 4446 CG2 THR A 589 −12.085 28.018−45.410 1.00 20.03 ATOM 4447 C THR A 589 −8.424 26.819 −45.427 1.0019.42 ATOM 4448 O THR A 589 −7.694 27.767 −45.743 1.00 18.66 ATOM 4449 NGLN A 590 −7.995 25.772 −44.734 1.00 19.62 ATOM 4450 CA GLN A 590 −6.60625.629 −44.317 1.00 20.20 ATOM 4451 CB GLN A 590 −6.359 26.261 −42.9391.00 21.06 ATOM 4452 CG GLN A 590 −4.950 25.956 −42.410 1.00 27.00 ATOM4453 CD GLN A 590 −4.184 27.189 −41.989 1.00 33.74 ATOM 4454 OE1 GLN A590 −4.771 28.196 −41.611 1.00 37.22 ATOM 4455 NE2 GLN A 590 −2.85527.118 −42.066 1.00 36.77 ATOM 4456 C GLN A 590 −6.247 24.159 −44.2951.00 19.00 ATOM 4457 O GLN A 590 −7.004 23.335 −43.771 1.00 18.70 ATOM4458 N VAL A 591 −5.113 23.811 −44.904 1.00 17.70 ATOM 4459 CA VAL A 591−4.682 22.404 −44.940 1.00 17.15 ATOM 4460 CB VAL A 591 −4.843 21.750−46.330 1.00 17.82 ATOM 4461 CG1 VAL A 591 −6.316 21.701 −46.744 1.0018.11 ATOM 4462 CG2 VAL A 591 −3.970 22.470 −47.390 1.00 17.60 ATOM 4463C VAL A 591 −3.213 22.360 −44.551 1.00 17.42 ATOM 4464 O VAL A 591−2.531 23.377 −44.638 1.00 17.32 ATOM 4465 N VAL A 592 −2.731 21.206−44.090 1.00 17.26 ATOM 4466 CA VAL A 592 −1.291 21.092 −43.887 1.0017.03 ATOM 4467 CB VAL A 592 −0.762 21.198 −42.365 1.00 18.70 ATOM 4468CG1 VAL A 592 0.335 20.217 −41.930 1.00 18.75 ATOM 4469 CG2 VAL A 592−1.810 21.731 −41.315 1.00 15.36 ATOM 4470 C VAL A 592 −0.736 19.951−44.730 1.00 16.99 ATOM 4471 O VAL A 592 −1.318 18.862 −44.828 1.0016.23 ATOM 4472 N LYS A 593 0.357 20.253 −45.403 1.00 15.38 ATOM 4473 CALYS A 593 0.953 19.302 −46.293 1.00 16.27 ATOM 4474 CB LYS A 593 1.30120.010 −47.616 1.00 16.69 ATOM 4475 CG LYS A 593 1.835 19.096 −48.6941.00 20.55 ATOM 4476 CD LYS A 593 0.791 18.101 −49.203 1.00 24.73 ATOM4477 CE LYS A 593 1.330 17.311 −50.409 1.00 27.26 ATOM 4478 NZ LYS A 5932.395 16.299 −50.074 1.00 28.37 ATOM 4479 C LYS A 593 2.209 18.783−45.588 1.00 16.01 ATOM 4480 O LYS A 593 3.175 19.525 −45.427 1.00 15.12ATOM 4481 N GLU A 594 2.195 17.519 −45.175 1.00 15.82 ATOM 4482 CA GLU A594 3.308 16.969 −44.407 1.00 16.23 ATOM 4483 CB GLU A 594 2.798 16.017−43.317 1.00 16.26 ATOM 4484 CG GLU A 594 1.866 16.732 −42.299 1.0016.62 ATOM 4485 CD GLU A 594 1.727 15.949 −40.991 1.00 18.94 ATOM 4486OE1 GLU A 594 1.267 14.778 −41.024 1.00 21.31 ATOM 4487 OE2 GLU A 5942.107 16.507 −39.940 1.00 16.43 ATOM 4488 C GLU A 594 4.286 16.245−45.323 1.00 17.34 ATOM 4489 O GLU A 594 3.973 15.177 −45.852 1.00 17.78ATOM 4490 N ASP A 595 5.463 16.833 −45.487 1.00 17.05 ATOM 4491 CA ASP A595 6.481 16.326 −46.405 1.00 17.21 ATOM 4492 CB ASP A 595 6.823 17.379−47.475 1.00 16.63 ATOM 4493 CG ASP A 595 5.678 17.619 −48.455 1.0017.91 ATOM 4494 OD1 ASP A 595 5.023 16.631 −48.857 1.00 20.73 ATOM 4495OD2 ASP A 595 5.434 18.795 −48.844 1.00 18.08 ATOM 4496 C ASP A 5957.734 15.955 −45.631 1.00 17.47 ATOM 4497 O ASP A 595 7.915 16.375−44.492 1.00 16.44 ATOM 4498 N THR A 596 8.598 15.162 −46.277 1.00 18.11ATOM 4499 CA THR A 596 9.917 14.835 −45.747 1.00 18.85 ATOM 4500 CB THRA 596 9.991 13.390 −45.188 1.00 19.58 ATOM 4501 OG1 THR A 596 9.05713.248 −44.116 1.00 20.97 ATOM 4502 CG2 THR A 596 11.385 13.124 −44.5981.00 20.94 ATOM 4503 C THR A 596 10.895 14.978 −46.914 1.00 19.34 ATOM4504 O THR A 596 10.588 14.531 −48.024 1.00 19.05 ATOM 4505 N TRP A 59712.050 15.581 −46.631 1.00 20.24 ATOM 4506 CA TRP A 597 13.074 15.921−47.633 1.00 21.94 ATOM 4507 CB TRP A 597 14.325 16.453 −46.940 1.0022.34 ATOM 4508 CG TRP A 597 15.445 16.854 −47.882 1.00 23.75 ATOM 4509CD1 TRP A 597 16.509 16.079 −48.275 1.00 25.08 ATOM 4510 NE1 TRP A 59717.327 16.801 −49.138 1.00 24.85 ATOM 4511 CE2 TRP A 597 16.802 18.059−49.300 1.00 25.25 ATOM 4512 CD2 TRP A 597 15.611 18.128 −48.527 1.0024.09 ATOM 4513 CE3 TRP A 597 14.875 19.325 −48.519 1.00 24.91 ATOM 4514CZ3 TRP A 597 15.334 20.401 −49.262 1.00 23.38 ATOM 4515 CH2 TRP A 59716.520 20.299 −50.028 1.00 25.14 ATOM 4516 CZ2 TRP A 597 17.265 19.137−50.053 1.00 22.49 ATOM 4517 C TRP A 597 13.424 14.708 −48.473 1.0023.42 ATOM 4518 O TRP A 597 13.675 13.635 −47.939 1.00 22.37 ATOM 4519 NGLN A 598 13.409 14.904 −49.788 1.00 25.26 ATOM 4520 CA GLN A 598 13.69813.850 −50.755 1.00 27.05 ATOM 4521 CB GLN A 598 12.936 14.124 −52.0521.00 26.51 ATOM 4522 CG GLN A 598 11.418 13.948 −51.895 1.00 26.10 ATOM4523 CD GLN A 598 10.642 14.209 −53.156 1.00 26.76 ATOM 4524 OE1 GLN A598 11.194 14.620 −54.175 1.00 27.68 ATOM 4525 NE2 GLN A 598 9.34013.990 −53.095 1.00 25.96 ATOM 4526 C GLN A 598 15.204 13.787 −50.9771.00 29.21 ATOM 4527 O GLN A 598 15.794 14.694 −51.574 1.00 28.61 ATOM4528 N SER A 599 15.818 12.722 −50.453 1.00 32.41 ATOM 4529 CA SER A 59917.273 12.530 −50.498 1.00 35.70 ATOM 4530 CB SER A 599 17.747 11.698−49.302 1.00 35.61 ATOM 4531 OG SER A 599 17.374 12.296 −48.072 1.0039.62 ATOM 4532 C SER A 599 17.703 11.831 −51.785 1.00 36.66 ATOM 4533 OSER A 599 16.916 11.145 −52.433 1.00 37.44 ATOM 4534 OXT SER A 59918.863 11.922 −52.194 1.00 38.18 ATOM 4535 C1 MAN A 601 −3.602 −3.018−46.412 1.00 102.64 ATOM 4536 C2 MAN A 601 −4.584 −2.109 −47.156 1.00102.73 ATOM 4537 O2 MAN A 601 −3.951 −1.548 −48.288 1.00 102.91 ATOM4538 C3 MAN A 601 −5.867 −2.845 −47.570 1.00 102.38 ATOM 4539 O3 MAN A601 −6.544 −2.112 −48.566 1.00 102.32 ATOM 4540 C4 MAN A 601 −5.640−4.269 −48.082 1.00 102.18 ATOM 4541 O4 MAN A 601 −6.860 −4.967 −47.9841.00 101.76 ATOM 4542 C5 MAN A 601 −4.561 −5.018 −47.298 1.00 102.40ATOM 4543 C6 MAN A 601 −4.172 −6.307 −48.010 1.00 102.48 ATOM 4544 O6MAN A 601 −3.156 −6.957 −47.280 1.00 102.80 ATOM 4545 O5 MAN A 601−3.400 −4.222 −47.131 1.00 102.71 ATOM 4546 C1 MAN A 602 −29.428 −4.974−42.477 1.00 77.32 ATOM 4547 C2 MAN A 602 −28.973 −6.434 −42.405 1.0077.44 ATOM 4548 O2 MAN A 602 −30.120 −7.253 −42.347 1.00 77.72 ATOM 4549C3 MAN A 602 −28.044 −6.835 −43.565 1.00 77.20 ATOM 4550 O3 MAN A 602−27.940 −8.239 −43.664 1.00 76.96 ATOM 4551 C4 MAN A 602 −28.487 −6.260−44.909 1.00 77.22 ATOM 4552 O4 MAN A 602 −27.471 −6.474 −45.862 1.0076.90 ATOM 4553 C5 MAN A 602 −28.766 −4.766 −44.768 1.00 77.47 ATOM 4554C6 MAN A 602 −29.185 −4.115 −46.081 1.00 77.84 ATOM 4555 O6 MAN A 602−28.163 −3.228 −46.483 1.00 78.24 ATOM 4556 O5 MAN A 602 −29.768 −4.562−43.790 1.00 77.46 ATOM 4557 C1 MAN A 603 −18.689 25.235 −53.677 1.0047.04 ATOM 4558 C2 MAN A 603 −20.074 24.872 −53.114 1.00 51.09 ATOM 4559O2 MAN A 603 −21.044 25.065 −54.120 1.00 52.42 ATOM 4560 C3 MAN A 603−20.141 23.420 −52.620 1.00 51.78 ATOM 4561 O3 MAN A 603 −21.465 23.079−52.262 1.00 53.36 ATOM 4562 C4 MAN A 603 −19.602 22.466 −53.686 1.0051.72 ATOM 4563 O4 MAN A 603 −19.615 21.142 −53.209 1.00 51.94 ATOM 4564C5 MAN A 603 −18.179 22.911 −54.021 1.00 51.66 ATOM 4565 C6 MAN A 603−17.421 21.906 −54.892 1.00 53.80 ATOM 4566 O6 MAN A 603 −17.915 21.885−56.214 1.00 55.49 ATOM 4567 O5 MAN A 603 −18.217 24.223 −54.581 1.0049.62 ATOM 4568 C1 MAN A 605 −4.678 15.117 −57.896 1.00 58.79 ATOM 4569C2 MAN A 605 −3.360 15.555 −58.538 1.00 58.65 ATOM 4570 O2 MAN A 605−2.564 14.412 −58.722 1.00 59.38 ATOM 4571 C3 MAN A 605 −3.570 16.269−59.878 1.00 58.70 ATOM 4572 O3 MAN A 605 −2.523 15.985 −60.778 1.0059.27 ATOM 4573 C4 MAN A 605 −4.915 15.892 −60.491 1.00 58.73 ATOM 4574O4 MAN A 605 −5.084 16.538 −61.730 1.00 59.62 ATOM 4575 C5 MAN A 605−6.054 16.284 −59.547 1.00 58.90 ATOM 4576 C6 MAN A 605 −7.370 15.612−59.932 1.00 58.75 ATOM 4577 O6 MAN A 605 −7.255 14.219 −59.738 1.0059.42 ATOM 4578 O5 MAN A 605 −5.730 16.034 −58.173 1.00 58.30 ATOM 4579C1 MAN A 606 −10.273 28.688 −42.727 1.00 25.90 ATOM 4580 C2 MAN A 606−9.839 27.944 −41.452 1.00 28.64 ATOM 4581 O2 MAN A 606 −9.245 28.909−40.620 1.00 28.84 ATOM 4582 C3 MAN A 606 −10.999 27.249 −40.710 1.0029.65 ATOM 4583 O3 MAN A 606 −10.568 26.763 −39.441 1.00 28.85 ATOM 4584C4 MAN A 606 −12.203 28.177 −40.551 1.00 30.36 ATOM 4585 O4 MAN A 606−13.330 27.463 −40.084 1.00 30.29 ATOM 4586 C5 MAN A 606 −12.553 28.769−41.914 1.00 30.72 ATOM 4587 C6 MAN A 606 −13.730 29.731 −41.778 1.0033.97 ATOM 4588 O6 MAN A 606 −13.624 30.732 −42.762 1.00 36.82 ATOM 4589O5 MAN A 606 −11.434 29.464 −42.435 1.00 28.12 ATOM 4590 C1 MAN A 607−31.396 1.963 −40.521 1.00 50.29 ATOM 4591 C2 MAN A 607 −30.220 1.790−41.485 1.00 52.65 ATOM 4592 O2 MAN A 607 −30.541 0.785 −42.419 1.0054.93 ATOM 4593 C3 MAN A 607 −29.845 3.092 −42.208 1.00 52.48 ATOM 4594O3 MAN A 607 −28.932 2.836 −43.251 1.00 53.01 ATOM 4595 C4 MAN A 607−31.068 3.818 −42.766 1.00 52.78 ATOM 4596 O4 MAN A 607 −30.672 5.070−43.297 1.00 52.92 ATOM 4597 C5 MAN A 607 −32.103 3.985 −41.652 1.0052.23 ATOM 4598 C6 MAN A 607 −33.331 4.749 −42.153 1.00 52.96 ATOM 4599O6 MAN A 607 −34.520 4.076 −41.791 1.00 52.95 ATOM 4600 O5 MAN A 607−32.451 2.702 −41.127 1.00 51.79 ATOM 4601 C1 MAN A 608 3.870 15.416−59.489 1.00 37.21 ATOM 4602 C2 MAN A 608 5.134 15.938 −60.168 1.0040.45 ATOM 4603 O2 MAN A 608 6.091 14.903 −60.120 1.00 38.47 ATOM 4604C3 MAN A 608 4.872 16.381 −61.608 1.00 42.66 ATOM 4605 O3 MAN A 6086.071 16.726 −62.263 1.00 44.20 ATOM 4606 C4 MAN A 608 4.122 15.321−62.401 1.00 44.80 ATOM 4607 O4 MAN A 608 3.708 15.907 −63.612 1.0047.73 ATOM 4608 C5 MAN A 608 2.893 14.887 −61.597 1.00 44.80 ATOM 4609C6 MAN A 608 2.042 13.861 −62.342 1.00 47.55 ATOM 4610 O6 MAN A 6081.085 14.582 −63.104 1.00 49.87 ATOM 4611 O5 MAN A 608 3.262 14.423−60.302 1.00 42.18 ATOM 4612 C1 NAG A 611 3.450 −2.354 −8.282 1.00 23.44ATOM 4613 C2 NAG A 611 3.474 −0.875 −7.878 1.00 24.51 ATOM 4614 N2 NAG A611 4.425 −0.077 −8.630 1.00 21.95 ATOM 4615 C7 NAG A 611 4.123 0.454−9.818 1.00 22.94 ATOM 4616 O7 NAG A 611 3.030 0.322 −10.367 1.00 20.93ATOM 4617 C8 NAG A 611 5.216 1.232 −10.481 1.00 21.54 ATOM 4618 C3 NAG A611 3.741 −0.713 −6.380 1.00 25.60 ATOM 4619 O3 NAG A 611 3.676 0.655−6.047 1.00 24.91 ATOM 4620 C4 NAG A 611 2.741 −1.528 −5.554 1.00 25.70ATOM 4621 O4 NAG A 611 3.196 −1.598 −4.227 1.00 28.27 ATOM 4622 C5 NAG A611 2.648 −2.952 −6.086 1.00 26.18 ATOM 4623 C6 NAG A 611 1.524 −3.738−5.397 1.00 26.64 ATOM 4624 O6 NAG A 611 0.278 −3.081 −5.497 1.00 25.38ATOM 4625 O5 NAG A 611 2.437 −2.975 −7.488 1.00 24.34 ATOM 4626 C1 NAG A612 2.499 −0.713 −3.326 1.00 32.04 ATOM 4627 C2 NAG A 612 2.710 −1.192−1.879 1.00 35.81 ATOM 4628 N2 NAG A 612 2.254 −2.556 −1.666 1.00 37.89ATOM 4629 C7 NAG A 612 3.072 −3.605 −1.753 1.00 39.19 ATOM 4630 O7 NAG A612 4.277 −3.517 −2.031 1.00 40.58 ATOM 4631 C8 NAG A 612 2.439 −4.947−1.507 1.00 38.98 ATOM 4632 C3 NAG A 612 2.012 −0.256 −0.899 1.00 37.96ATOM 4633 O3 NAG A 612 2.352 −0.666 0.403 1.00 41.23 ATOM 4634 C4 NAG A612 2.491 1.176 −1.129 1.00 37.63 ATOM 4635 O4 NAG A 612 1.789 2.053−0.278 1.00 40.85 ATOM 4636 C5 NAG A 612 2.294 1.565 −2.604 1.00 35.10ATOM 4637 C6 NAG A 612 2.785 2.982 −2.903 1.00 31.93 ATOM 4638 O6 NAG A612 4.188 2.994 −3.008 0.58 32.70 ATOM 4639 O5 NAG A 612 2.974 0.625−3.425 1.00 31.95 ATOM 4640 O8 BTB A 620 −1.213 18.638 −21.639 1.0023.78 ATOM 4641 C8 BTB A 620 −1.255 19.440 −22.838 1.00 17.50 ATOM 4642C7 BTB A 620 −2.257 18.851 −23.831 1.00 15.39 ATOM 4643 N BTB A 620−1.808 17.505 −24.294 1.00 13.88 ATOM 4644 C5 BTB A 620 −1.274 17.600−25.684 1.00 12.99 ATOM 4645 C6 BTB A 620 0.017 18.399 −25.786 1.0014.67 ATOM 4646 O6 BTB A 620 0.949 18.004 −24.768 1.00 16.93 ATOM 4647C2 BTB A 620 −2.926 16.495 −24.191 1.00 13.33 ATOM 4648 C4 BTB A 620−4.238 16.972 −24.835 1.00 13.45 ATOM 4649 O4 BTB A 620 −4.167 17.018−26.265 1.00 14.77 ATOM 4650 C3 BTB A 620 −3.213 16.295 −22.703 1.0013.18 ATOM 4651 O3 BTB A 620 −1.984 15.920 −22.059 1.00 12.74 ATOM 4652C1 BTB A 620 −2.501 15.161 −24.845 1.00 13.57 ATOM 4653 O1 BTB A 620−3.463 14.138 −24.525 1.00 13.07 ATOM 4654 O WAT W 1 −7.741 16.530−28.587 1.00 12.90 ATOM 4655 O WAT W 2 −1.955 18.721 −7.814 1.00 11.77ATOM 4656 O WAT W 3 −17.101 16.033 −19.836 1.00 15.26 ATOM 4657 O WAT W4 −1.389 7.464 −24.070 1.00 15.86 ATOM 4658 O WAT W 5 −8.070 20.758−43.462 1.00 19.56 ATOM 4659 O WAT W 6 −12.959 28.534 −26.860 1.00 16.12ATOM 4660 O WAT W 7 −0.502 31.488 −57.004 1.00 33.06 ATOM 4661 O WAT W 82.095 5.710 −17.808 1.00 18.68 ATOM 4662 O WAT W 9 −7.601 14.567 −6.8271.00 14.97 ATOM 4663 O WAT W 10 24.863 23.325 −37.431 1.00 32.31 ATOM4664 O WAT W 11 −22.569 7.289 −10.357 1.00 17.52 ATOM 4665 O WAT W 12−18.987 1.758 −22.078 1.00 23.03 ATOM 4666 O WAT W 13 −3.226 16.264−54.338 1.00 32.98 ATOM 4667 O WAT W 14 6.141 16.546 −42.196 1.00 16.00ATOM 4668 O WAT W 15 −10.356 21.827 −22.675 1.00 13.52 ATOM 4669 O WAT W16 −3.130 25.355 −17.925 1.00 14.01 ATOM 4670 O WAT W 17 −11.823 29.479−29.411 1.00 17.50 ATOM 4671 O WAT W 18 −14.383 15.964 −19.553 1.0013.02 ATOM 4672 O WAT W 19 −1.180 16.935 −10.101 1.00 18.86 ATOM 4673 OWAT W 20 −31.133 23.501 4.462 1.00 16.66 ATOM 4674 O WAT W 21 −4.81924.193 −15.023 1.00 14.31 ATOM 4675 O WAT W 22 1.709 22.276 −4.126 1.0021.96 ATOM 4676 O WAT W 23 −5.339 21.386 −7.463 1.00 15.78 ATOM 4677 OWAT W 24 −17.232 15.476 1.374 1.00 17.64 ATOM 4678 O WAT W 25 −11.4494.860 −24.929 1.00 17.45 ATOM 4679 O WAT W 26 −17.555 17.679 −39.8151.00 23.23 ATOM 4680 O WAT W 27 10.075 17.015 −49.295 1.00 24.05 ATOM4681 O WAT W 28 −16.018 −0.740 −24.205 1.00 18.07 ATOM 4682 O WAT W 299.446 24.991 −37.612 1.00 19.20 ATOM 4683 O WAT W 30 −4.165 26.137−12.642 1.00 18.33 ATOM 4684 O WAT W 31 2.771 22.947 −14.916 1.00 25.80ATOM 4685 O WAT W 32 −12.297 21.394 −35.680 1.00 14.89 ATOM 4686 O WAT W33 −24.061 13.570 10.081 1.00 24.96 ATOM 4687 O WAT W 34 10.032 29.725−56.684 1.00 26.97 ATOM 4688 O WAT W 35 0.231 4.133 −28.595 1.00 17.67ATOM 4689 O WAT W 36 0.335 2.173 −30.650 1.00 18.32 ATOM 4690 O WAT W 37−10.199 24.315 −42.717 1.00 22.38 ATOM 4691 O WAT W 38 −14.151 12.872−8.204 1.00 16.16 ATOM 4692 O WAT W 39 −2.710 9.564 −16.092 1.00 14.10ATOM 4693 O WAT W 40 5.954 7.990 −32.401 1.00 16.59 ATOM 4694 O WAT W 410.294 5.561 −25.249 1.00 17.87 ATOM 4695 O WAT W 42 2.102 15.148 −37.7181.00 14.64 ATOM 4696 O WAT W 43 −19.351 1.384 −26.295 1.00 20.27 ATOM4697 O WAT W 44 −19.623 9.533 −17.751 1.00 14.67 ATOM 4698 O WAT W 453.117 18.767 −36.336 1.00 12.66 ATOM 4699 O WAT W 46 −15.016 16.9500.662 1.00 20.14 ATOM 4700 O WAT W 47 −22.261 4.600 −10.993 1.00 16.38ATOM 4701 O WAT W 48 −12.926 5.474 −22.680 1.00 19.85 ATOM 4702 O WAT W49 5.564 17.071 −37.018 1.00 16.82 ATOM 4703 O WAT W 50 −19.848 20.552−2.718 1.00 19.38 ATOM 4704 O WAT W 51 −15.859 17.744 −41.901 1.00 19.69ATOM 4705 O WAT W 52 −16.430 25.522 −1.123 1.00 19.97 ATOM 4706 O WAT W53 −15.978 5.366 −12.193 1.00 26.02 ATOM 4707 O WAT W 54 −1.637 9.365−26.035 1.00 14.42 ATOM 4708 O WAT W 55 −10.759 27.212 −30.898 1.0017.28 ATOM 4709 O WAT W 56 −11.509 0.756 −13.101 1.00 20.70 ATOM 4710 OWAT W 57 −16.950 15.108 4.727 1.00 23.86 ATOM 4711 O WAT W 58 −25.36826.009 −7.106 1.00 25.08 ATOM 4712 O WAT W 59 −16.870 22.937 −3.651 1.0017.56 ATOM 4713 O WAT W 60 −14.388 13.258 −40.897 1.00 27.90 ATOM 4714 OWAT W 61 −1.509 −4.779 −6.723 1.00 31.01 ATOM 4715 O WAT W 62 −1.97327.723 −11.521 1.00 21.99 ATOM 4716 O WAT W 63 −1.159 −10.623 −29.5921.00 36.68 ATOM 4717 O WAT W 64 −1.943 16.930 −42.957 1.00 21.57 ATOM4718 O WAT W 65 −1.507 25.238 −40.032 1.00 31.36 ATOM 4719 O WAT W 66−4.023 5.499 −31.787 1.00 20.03 ATOM 4720 O WAT W 67 −13.383 13.873−21.065 1.00 12.01 ATOM 4721 O WAT W 68 −15.098 10.726 −24.467 1.0024.34 ATOM 4722 O WAT W 69 −2.122 13.975 −13.435 1.00 12.51 ATOM 4723 OWAT W 70 −4.807 19.360 −43.270 1.00 20.93 ATOM 4724 O WAT W 71 −26.02826.143 −33.768 1.00 28.78 ATOM 4725 O WAT W 72 −19.347 21.638 3.482 1.0019.92 ATOM 4726 O WAT W 73 −27.299 24.219 −6.045 1.00 20.97 ATOM 4727 OWAT W 74 −21.114 −0.343 −28.050 1.00 22.38 ATOM 4728 O WAT W 75 −5.81834.483 −11.645 1.00 20.61 ATOM 4729 O WAT W 76 6.048 1.098 −23.393 1.0016.77 ATOM 4730 O WAT W 77 −3.946 23.711 −39.552 1.00 25.07 ATOM 4731 OWAT W 78 −18.572 21.631 −41.884 1.00 25.98 ATOM 4732 O WAT W 79 5.23926.273 −31.646 1.00 27.95 ATOM 4733 O WAT W 80 0.054 15.597 −45.905 1.0028.45 ATOM 4734 O WAT W 81 −3.130 21.534 −5.652 1.00 20.95 ATOM 4735 OWAT W 82 −12.534 4.331 −20.095 1.00 17.49 ATOM 4736 O WAT W 83 0.78516.541 −14.558 1.00 14.65 ATOM 4737 O WAT W 84 −5.197 12.827 −31.5531.00 14.10 ATOM 4738 O WAT W 85 −16.738 26.994 −34.463 1.00 23.74 ATOM4739 O WAT W 86 3.596 22.076 −36.828 1.00 22.68 ATOM 4740 O WAT W 875.170 14.460 −40.572 1.00 26.56 ATOM 4741 O WAT W 88 −12.322 21.0500.328 1.00 30.36 ATOM 4742 O WAT W 89 7.426 14.327 −48.857 1.00 26.44ATOM 4743 O WAT W 90 −13.702 19.025 1.863 1.00 28.28 ATOM 4744 O WAT W91 8.794 2.010 −23.444 1.00 34.69 ATOM 4745 O WAT W 92 −6.185 5.529−30.210 1.00 17.03 ATOM 4746 O WAT W 93 −18.081 20.709 −4.839 1.00 17.61ATOM 4747 O WAT W 94 −15.469 13.082 −22.717 1.00 15.55 ATOM 4748 O WAT W95 13.101 16.811 −29.771 1.00 29.16 ATOM 4749 O WAT W 96 −25.944 7.031−2.628 1.00 27.42 ATOM 4750 O WAT W 97 −4.552 34.207 −7.388 1.00 22.19ATOM 4751 O WAT W 98 −2.231 −9.858 −32.291 1.00 26.28 ATOM 4752 O WAT W99 5.314 10.271 −28.762 1.00 29.39 ATOM 4753 O WAT W 100 −15.379 27.478−46.620 1.00 37.77 ATOM 4754 O WAT W 101 26.815 24.874 −36.295 1.0031.11 ATOM 4755 O WAT W 102 −18.489 −0.112 −24.256 1.00 23.63 ATOM 4756O WAT W 103 −23.763 26.890 −9.454 1.00 20.50 ATOM 4757 O WAT W 104−10.933 23.904 −50.315 1.00 25.63 ATOM 4758 O WAT W 105 5.864 12.071−41.668 1.00 29.27 ATOM 4759 O WAT W 106 2.526 9.409 −13.116 1.00 20.75ATOM 4760 O WAT W 107 −11.557 −6.653 −10.981 1.00 30.68 ATOM 4761 O WATW 108 −14.882 7.238 −22.254 1.00 24.81 ATOM 4762 O WAT W 109 −5.331−13.390 −25.293 1.00 35.63 ATOM 4763 O WAT W 110 −8.068 24.248 −40.5341.00 39.14 ATOM 4764 O WAT W 111 −0.779 14.419 −43.060 1.00 24.01 ATOM4765 O WAT W 112 −22.279 12.054 −26.750 1.00 32.13 ATOM 4766 O WAT W 113−26.829 1.352 −33.787 1.00 26.97 ATOM 4767 O WAT W 114 −14.120 14.1163.214 1.00 36.89 ATOM 4768 O WAT W 115 0.582 −9.914 −21.103 1.00 23.30ATOM 4769 O WAT W 116 −24.305 22.723 6.995 1.00 21.67 ATOM 4770 O WAT W117 −28.275 12.468 −15.419 1.00 21.87 ATOM 4771 O WAT W 118 3.699 27.669−20.781 1.00 34.08 ATOM 4772 O WAT W 119 −30.428 26.452 3.757 1.00 24.51ATOM 4773 O WAT W 120 19.168 26.858 −59.022 1.00 33.45 ATOM 4774 O WAT W121 −8.803 21.729 −1.693 1.00 18.30 ATOM 4775 O WAT W 122 2.863 1.621−31.755 1.00 19.96 ATOM 4776 O WAT W 123 −2.357 28.930 −56.725 1.0035.51 ATOM 4777 O WAT W 124 −16.780 5.504 −21.523 1.00 31.34 ATOM 4778 OWAT W 125 6.216 18.141 −30.592 1.00 20.87 ATOM 4779 O WAT W 126 11.78932.722 −38.773 1.00 40.41 ATOM 4780 O WAT W 127 −5.001 7.195 −45.6561.00 35.03 ATOM 4781 O WAT W 128 −18.743 1.608 −1.861 1.00 32.49 ATOM4782 O WAT W 129 −25.089 −1.945 −20.935 1.00 35.17 ATOM 4783 O WAT W 130−7.097 −2.177 −28.928 1.00 30.93 ATOM 4784 O WAT W 131 −12.591 2.907−11.929 1.00 18.60 ATOM 4785 O WAT W 132 −17.913 −2.374 −39.429 1.0029.36 ATOM 4786 O WAT W 133 −6.507 −7.038 −37.710 1.00 37.27 ATOM 4787 OWAT W 134 −0.628 7.596 −18.660 1.00 20.01 ATOM 4788 O WAT W 135 −11.68328.527 −37.016 1.00 36.45 ATOM 4789 O WAT W 136 −3.169 33.267 −18.0491.00 24.89 ATOM 4790 O WAT W 137 −16.742 8.938 −23.161 1.00 26.79 ATOM4791 O WAT W 138 −28.456 17.726 −22.449 1.00 32.61 ATOM 4792 O WAT W 13925.559 27.237 −45.392 1.00 43.21 ATOM 4793 O WAT W 140 −26.925 5.789−41.722 1.00 26.97 ATOM 4794 O WAT W 141 −16.907 20.013 −43.283 1.0029.68 ATOM 4795 O WAT W 142 −20.029 5.119 −1.799 1.00 24.76 ATOM 4796 OWAT W 143 8.706 1.050 −13.115 1.00 32.81 ATOM 4797 O WAT W 144 −4.35322.506 −1.252 1.00 24.86 ATOM 4798 O WAT W 145 −29.660 14.750 −15.2951.00 32.62 ATOM 4799 O WAT W 146 2.173 2.240 −12.124 1.00 26.38 ATOM4800 O WAT W 147 4.174 −4.659 −14.794 1.00 25.20 ATOM 4801 O WAT W 148−10.913 29.083 −33.130 1.00 27.78 ATOM 4802 O WAT W 149 −21.448 30.157−10.670 1.00 25.07 ATOM 4803 O WAT W 150 −23.296 18.641 −36.646 1.0027.11 ATOM 4804 O WAT W 151 −19.426 8.262 −24.240 1.00 25.11 ATOM 4805 OWAT W 152 4.729 −0.512 −31.679 1.00 23.50 ATOM 4806 O WAT W 153 9.24719.703 −33.306 1.00 23.44 ATOM 4807 O WAT W 154 6.024 15.401 −22.7681.00 27.11 ATOM 4808 O WAT W 155 −16.077 30.180 −4.530 1.00 23.52 ATOM4809 O WAT W 156 −0.038 14.751 −8.812 1.00 25.64 ATOM 4810 O WAT W 1572.962 18.631 −29.190 1.00 18.13 ATOM 4811 O WAT W 158 8.793 12.371−36.745 1.00 23.77 ATOM 4812 O WAT W 159 −22.406 9.468 −0.415 1.00 21.66ATOM 4813 O WAT W 160 −10.961 33.685 −7.076 1.00 25.30 ATOM 4814 O WAT W161 −8.504 27.891 −3.964 1.00 33.88 ATOM 4815 O WAT W 162 6.836 20.663−32.439 1.00 24.97 ATOM 4816 O WAT W 163 4.292 23.232 −29.206 1.00 32.74ATOM 4817 O WAT W 164 2.350 3.656 −15.645 1.00 23.29 ATOM 4818 O WAT W165 −17.377 10.190 −20.605 1.00 25.21 ATOM 4819 O WAT W 166 −23.42624.714 4.551 1.00 26.12 ATOM 4820 O WAT W 167 0.338 1.730 −14.995 1.0031.29 ATOM 4821 O WAT W 168 −3.303 17.836 −46.350 1.00 28.34 ATOM 4822 OWAT W 169 1.465 6.514 −14.840 1.00 22.81 ATOM 4823 O WAT W 170 2.40911.466 −4.481 1.00 29.82 ATOM 4824 O WAT W 171 0.998 19.313 −20.348 1.0031.57 ATOM 4825 O WAT W 172 7.556 −3.076 −34.213 1.00 31.62 ATOM 4826 OWAT W 173 −25.163 1.132 −18.852 1.00 33.45 ATOM 4827 O WAT W 174 −25.60617.471 −26.509 1.00 27.89 ATOM 4828 O WAT W 175 5.952 32.621 −65.9551.00 42.20 ATOM 4829 O WAT W 176 −27.397 26.421 −12.489 1.00 29.06 ATOM4830 O WAT W 177 −17.506 35.918 −29.284 1.00 36.40 ATOM 4831 O WAT W 178−18.298 7.055 −19.628 1.00 30.04 ATOM 4832 O WAT W 179 −24.383 14.811−26.605 1.00 29.51 ATOM 4833 O WAT W 180 −1.204 27.462 −35.328 1.0029.93 ATOM 4834 O WAT W 181 −14.112 33.822 −23.916 1.00 34.66 ATOM 4835O WAT W 182 2.887 26.714 −9.619 1.00 34.18 ATOM 4836 O WAT W 183 −16.0624.698 1.046 1.00 32.44 ATOM 4837 O WAT W 184 −13.340 36.111 −4.359 1.0039.14 ATOM 4838 O WAT W 185 9.661 34.457 −47.977 1.00 37.66 ATOM 4839 OWAT W 186 −8.465 24.284 −1.237 1.00 33.71 ATOM 4840 O WAT W 187 16.97115.520 −43.951 1.00 42.49 ATOM 4841 O WAT W 188 −12.038 −14.614 −20.2991.00 34.37 ATOM 4842 O WAT W 189 −5.887 22.387 −40.784 1.00 33.70 ATOM4843 O WAT W 190 −3.962 −18.100 −17.720 1.00 31.33 ATOM 4844 O WAT W 191−30.888 11.643 −15.288 1.00 36.84 ATOM 4845 O WAT W 192 11.576 13.142−37.752 1.00 32.89 ATOM 4846 O WAT W 193 −7.856 3.348 −41.927 1.00 34.02ATOM 4847 O WAT W 194 −20.849 7.518 7.652 1.00 32.37 ATOM 4848 O WAT W195 16.954 13.938 −58.514 1.00 42.65 ATOM 4849 O WAT W 196 −31.884 7.593−13.893 1.00 37.54 ATOM 4850 O WAT W 197 4.560 −14.190 −17.137 1.0036.09 ATOM 4851 O WAT W 198 1.116 27.617 −39.051 1.00 37.08 ATOM 4852 OWAT W 199 −1.019 −12.134 −21.800 1.00 36.12 ATOM 4853 O WAT W 200 8.3500.111 −21.198 1.00 36.56 ATOM 4854 O WAT W 201 −2.691 31.235 −26.9101.00 32.08 ATOM 4855 O WAT W 202 13.222 30.530 −38.626 1.00 36.46 ATOM4856 O WAT W 203 −11.218 19.535 −54.549 1.00 35.12 ATOM 4857 O WAT W 204−5.623 10.865 −46.910 1.00 35.48 ATOM 4858 O WAT W 205 −18.073 1.743−43.946 1.00 40.62 ATOM 4859 O WAT W 206 −32.195 23.231 2.102 1.00 34.73ATOM 4860 O WAT W 207 −24.204 8.994 −2.941 1.00 30.29 ATOM 4861 O WAT W208 −4.771 18.292 −48.610 1.00 31.87 ATOM 4862 O WAT W 209 −17.15623.843 −40.674 1.00 35.73 ATOM 4863 O WAT W 210 8.319 13.422 −13.2971.00 37.43 ATOM 4864 O WAT W 211 −25.962 8.559 −33.791 1.00 33.46 ATOM4865 O WAT W 212 −36.129 8.276 3.147 1.00 40.24 ATOM 4866 O WAT W 21320.833 21.074 −56.185 1.00 39.11 ATOM 4867 O WAT W 214 −17.726 14.0878.330 1.00 39.10 ATOM 4868 O WAT W 215 8.944 8.011 −10.493 1.00 41.24ATOM 4869 O WAT W 216 −16.566 35.858 −11.282 1.00 38.90 ATOM 4870 O WATW 217 −20.560 11.198 −43.128 1.00 34.83 ATOM 4871 O WAT W 218 3.261−0.833 −39.177 1.00 32.67 ATOM 4872 O WAT W 219 −22.370 −13.152 −34.4121.00 59.42 ATOM 4873 O WAT W 220 −24.775 6.925 5.968 1.00 34.28 ATOM4874 O WAT W 221 −20.357 21.098 −45.702 1.00 36.83 ATOM 4875 O WAT W 2222.502 28.932 −40.686 1.00 36.85 ATOM 4876 O WAT W 223 −17.630 −5.533−21.334 1.00 35.08 ATOM 4877 O WAT W 224 −19.358 −1.912 −43.190 1.0035.83 ATOM 4878 O WAT W 225 −14.632 25.995 −42.094 1.00 41.12 ATOM 4879O WAT W 226 −28.967 5.606 −22.103 1.00 45.63 ATOM 4880 O WAT W 227−4.326 9.934 5.097 1.00 44.46 ATOM 4881 O WAT W 228 3.983 22.711 −17.3361.00 46.21 ATOM 4882 O WAT W 229 −17.238 16.931 6.949 1.00 42.44 ATOM4883 O WAT W 230 −25.871 18.809 10.364 1.00 36.15 ATOM 4884 O WAT W 231−23.524 31.294 −19.082 1.00 35.99 ATOM 4885 O WAT W 232 −5.261 −9.321−38.277 1.00 43.42 ATOM 4886 O WAT W 233 −22.757 28.188 −29.173 1.0039.56 ATOM 4887 O WAT W 234 −25.699 26.238 −0.299 1.00 40.26 ATOM 4888 OWAT W 235 −21.884 −4.007 −22.294 1.00 38.01 ATOM 4889 O WAT W 236 −6.696−17.346 −27.753 1.00 39.83 ATOM 4890 O WAT W 237 −18.052 5.515 3.8881.00 37.82 ATOM 4891 O WAT W 238 −6.073 35.708 −9.057 1.00 36.25 ATOM4892 O WAT W 239 −8.876 2.984 −44.368 1.00 45.85 ATOM 4893 O WAT W 2409.232 31.613 −65.496 1.00 45.82 ATOM 4894 O WAT W 241 −28.246 26.806−0.118 1.00 36.45 ATOM 4895 O WAT W 242 −27.793 14.675 −45.312 1.0052.10 ATOM 4896 O WAT W 243 7.463 14.079 −55.045 1.00 36.97 ATOM 4897 OWAT W 244 −28.572 4.769 −1.130 1.00 36.20 ATOM 4898 O WAT W 245 8.22112.936 −50.870 1.00 38.81 ATOM 4899 O WAT W 246 −23.302 −2.082 −27.1911.00 32.98 ATOM 4900 O WAT W 247 −13.035 8.248 −46.620 1.00 51.73 ATOM4901 O WAT W 248 −11.869 31.852 −50.157 1.00 56.13 ATOM 4902 O WAT W 2490.898 13.920 −6.521 1.00 27.38 ATOM 4903 O WAT W 250 20.427 30.852−45.446 1.00 35.54 ATOM 4904 O WAT W 251 −1.397 12.400 −44.617 1.0039.33 ATOM 4905 O WAT W 252 −27.354 24.696 −3.162 1.00 35.38 ATOM 4906 OWAT W 253 17.587 20.557 −31.069 1.00 41.51 ATOM 4907 O WAT W 254 −7.93635.055 −7.354 1.00 39.02 ATOM 4908 O WAT W 255 −22.469 7.215 −2.044 1.0038.25 ATOM 4909 O WAT W 256 2.038 15.474 −52.963 1.00 50.04 ATOM 4910 OWAT W 257 10.889 10.184 −21.700 1.00 44.84 ATOM 4911 O WAT W 258 −11.71410.583 4.136 1.00 42.70 ATOM 4912 O WAT W 259 −14.719 6.574 2.959 1.0043.18 ATOM 4913 O WAT W 260 −16.694 25.390 −37.688 1.00 36.77 ATOM 4914O WAT W 261 −9.212 13.388 −48.363 1.00 38.05 ATOM 4915 O WAT W 264−0.611 −1.965 −3.253 1.00 37.95 ATOM 4916 O WAT W 265 −16.380 30.998−14.262 1.00 32.44 ATOM 4917 O WAT W 266 9.420 16.012 −61.368 1.00 35.22ATOM 4918 O WAT W 267 −4.976 −15.180 −21.223 1.00 45.50 ATOM 4919 O WATW 268 −16.631 33.287 −14.201 1.00 34.50 ATOM 4920 O WAT W 269 −16.88334.052 −32.249 1.00 36.88 ATOM 4921 O WAT W 270 −8.293 −16.006 −14.5351.00 34.80 ATOM 4922 O WAT W 273 0.240 4.589 −13.868 1.00 32.34 ATOM4923 O WAT W 275 3.657 14.447 −55.516 1.00 43.54 ATOM 4924 O WAT W 276−17.602 20.784 −51.471 1.00 38.88 ATOM 4925 O WAT W 277 −10.479 31.683−30.513 1.00 40.35 ATOM 4926 O WAT W 278 −10.974 4.308 −5.745 1.00 40.30ATOM 4927 O WAT W 280 −4.336 36.908 −17.666 1.00 34.01 ATOM 4928 O WAT W281 6.720 33.970 −53.572 1.00 38.81 ATOM 4929 O WAT W 282 −30.457 23.527−0.621 1.00 34.61 ATOM 4930 O WAT W 283 16.969 17.394 −30.816 1.00 55.74ATOM 4931 O WAT W 284 −24.391 5.834 −24.909 1.00 37.92 ATOM 4932 O WAT W285 4.567 9.814 −4.438 1.00 44.83 ATOM 4933 O WAT W 286 −24.370 −7.328−27.875 1.00 56.18 ATOM 4934 O WAT W 287 −21.605 11.887 9.715 1.00 43.15ATOM 4935 O WAT W 288 8.603 0.412 −37.887 1.00 40.47 ATOM 4936 O WAT W290 −20.056 21.495 6.073 1.00 41.68 ATOM 4937 O WAT W 291 −3.221 28.158−33.448 1.00 44.24 ATOM 4938 O WAT W 292 9.171 9.103 −38.735 1.00 34.30ATOM 4939 O WAT W 293 2.894 22.763 −25.829 1.00 38.57 ATOM 4940 O WAT W294 −29.901 19.604 −14.929 1.00 37.66 ATOM 4941 O WAT W 296 −4.57930.229 −29.110 1.00 40.57 ATOM 4942 O WAT W 297 −23.821 11.441 −33.1871.00 42.81 ATOM 4943 O WAT W 298 −26.753 −3.087 −31.243 1.00 39.08 ATOM4944 O WAT W 300 −10.820 35.024 −53.050 1.00 55.39 ATOM 4945 O WAT W 302−1.992 7.169 −31.692 1.00 39.12 ATOM 4946 O WAT W 303 −15.282 −19.000−23.770 1.00 34.62 ATOM 4947 O WAT W 304 12.106 10.568 −25.112 1.0039.04 ATOM 4948 O WAT W 305 2.585 2.766 1.880 1.00 53.88 ATOM 4949 O WATW 306 3.680 21.122 −19.818 1.00 46.35 ATOM 4950 O WAT W 307 22.75924.721 −48.099 1.00 40.35 ATOM 4951 O WAT W 309 −17.062 −6.726 −19.2021.00 41.54 ATOM 4952 O WAT W 311 12.594 1.109 −31.461 1.00 47.85 ATOM4953 O WAT W 312 23.347 25.060 −50.638 1.00 49.64 ATOM 4954 O WAT W 314−18.291 4.422 −19.151 1.00 39.17 ATOM 4955 O WAT W 315 −11.815 −7.807−8.676 1.00 37.74 ATOM 4956 O WAT W 316 −25.147 1.885 −4.649 1.00 44.72ATOM 4957 O WAT W 317 −36.473 13.592 5.315 1.00 44.38 ATOM 4958 O WAT W318 −17.587 20.023 −46.231 1.00 48.30 ATOM 4959 O WAT W 319 −16.08129.024 −54.668 1.00 39.40 ATOM 4960 O WAT W 320 −14.210 32.143 −5.4941.00 42.73 ATOM 4961 O WAT W 321 −15.274 28.830 −38.916 1.00 46.47 ATOM4962 O WAT W 322 −32.792 22.221 −3.433 1.00 41.52 ATOM 4963 O WAT W 323−32.475 16.905 −12.401 1.00 46.29 ATOM 4964 O WAT W 325 15.341 22.212−60.490 1.00 34.67 ATOM 4965 O WAT W 326 −12.668 8.518 −41.723 1.0036.26 ATOM 4966 O WAT W 327 4.709 20.490 −10.568 1.00 38.04 ATOM 4967 OWAT W 328 13.937 10.625 −29.312 1.00 38.32 ATOM 4968 O WAT W 329 −21.9649.615 −24.896 1.00 40.43 ATOM 4969 O WAT W 330 19.325 25.925 −40.1991.00 51.36 ATOM 4970 O WAT W 331 −19.010 8.073 −45.255 1.00 46.06 ATOM4971 O WAT W 332 −25.024 −2.892 −29.306 1.00 41.74 ATOM 4972 O WAT W 333−16.593 −7.067 −23.297 1.00 38.54 ATOM 4973 O WAT W 334 −17.517 24.0782.157 1.00 45.82 ATOM 4974 O WAT W 335 −19.123 31.941 0.010 1.00 38.91ATOM 4975 O WAT W 337 10.677 21.901 −62.740 1.00 44.31 ATOM 4976 O WAT W338 4.510 15.230 −51.810 1.00 42.12 ATOM 4977 O WAT W 339 13.979 14.161−43.380 1.00 46.42 ATOM 4978 O WAT W 341 5.979 −11.625 −28.739 1.0043.09 ATOM 4979 O WAT W 342 −19.453 13.347 10.394 1.00 42.44 ATOM 4980 OWAT W 343 7.085 23.050 −30.796 1.00 34.94 ATOM 4981 O WAT W 345 6.47124.087 −63.943 1.00 45.16 ATOM 4982 O WAT W 347 3.734 22.842 −12.0311.00 43.85 ATOM 4983 O WAT W 348 −17.739 7.564 5.723 1.00 43.77 ATOM4984 O WAT W 351 −22.014 31.372 −24.708 1.00 42.69 ATOM 4985 O WAT W 35225.016 25.103 −46.967 1.00 40.51 ATOM 4986 O WAT W 353 7.969 32.464−67.637 1.00 57.78 ATOM 4987 O WAT W 354 −27.444 5.101 5.861 1.00 48.31ATOM 4988 O WAT W 356 8.012 11.087 −40.867 1.00 47.51 ATOM 4989 O WAT W357 4.974 29.116 −17.433 1.00 43.72 ATOM 4990 O WAT W 358 −0.457 9.488−45.288 1.00 45.63 ATOM 4991 O WAT W 360 −3.090 36.536 −12.138 1.0046.29 ATOM 4992 O WAT W 361 20.072 19.772 −36.896 1.00 38.73 ATOM 4993 OWAT W 363 −26.217 15.345 −28.735 1.00 49.07 ATOM 4994 O WAT W 365−25.308 0.100 −48.602 1.00 60.01 ATOM 4995 O WAT W 367 19.369 29.586−58.438 1.00 48.19 ATOM 4996 O WAT W 369 12.808 11.144 −54.427 1.0048.48 ATOM 4997 O WAT W 370 9.410 2.674 −16.115 1.00 44.59 ATOM 4998 OWAT W 372 −10.249 38.564 −13.215 1.00 48.60 ATOM 4999 O WAT W 373−24.151 16.211 10.617 1.00 42.16 ATOM 5000 O WAT W 375 −6.459 31.697−48.106 1.00 46.03 ATOM 5001 O WAT W 376 −11.605 27.116 −1.562 1.0044.58 ATOM 5002 O WAT W 377 −4.703 24.150 −62.673 1.00 48.70 ATOM 5003 OWAT W 379 6.889 0.036 −7.530 1.00 45.35 ATOM 5004 O WAT W 381 −13.60132.742 −32.002 1.00 51.37 ATOM 5005 O WAT W 383 −28.077 5.243 −4.6881.00 37.46 END

1. A glucoamylase variant comprising two or more amino acidsubstitutions corresponding to position 61, 73, 417, 430, 431, 503, 511,535, 539, or 563 of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase, wherein the glucoamylase variant has at least 80% sequenceidentity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or
 9. 2. The glucoamylasevariant of claim 1, wherein the parent glucoamylase has a catalyticdomain that has at least 80% sequence identity with SEQ ID NO: 1, 2, 3,5, 6, 7, 8, or 9, or a starch binding domain that has at least 80%sequence identity with SEQ ID NO: 1 or
 2. 3. The glucoamylase variant ofclaim 1, wherein the glucoamylase variant has at least 85% sequenceidentity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or
 9. 4. The glucoamylasevariant of claim 1, wherein the glucoamylase variant has at least 90%sequence identity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or
 9. 5. Theglucoamylase variant of claim 1, wherein the glucoamylase variant has atleast 95% sequence identity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9.6. The glucoamylase variant of claim 1, wherein the glucoamylase varianthas at least 99.5% sequence identity with SEQ ID NO: 1, 2, 3, 5, 6, 7,8, or
 9. 7. The glucoamylase variant of claim 1, wherein the parentglucoamylase is SEQ ID NO: 1 or
 2. 8. The glucoamylase variant of claim1, wherein the glucoamylase variant further comprises one or more aminoacid substitutions corresponding to position: 4, 5, 12, 24, 43, 44, 45,46, 47, 49, 51, 70, 75, 6, 94, 100, 108, 114, 116, 119, 122, 124, 125,137, 141, 143, 146, 148, 169, 171, 172, 175, 178, 180, 181, 208, 211,228, 242, 243, 245, 292, 294, 197, 309, 310, 313, 314, 315, 316, 317,321, 340, 341, 350, 353, 356, 363, 368, 369, 375, 376, 395, 398, 401,408, 409, 412, 415, 418, 421, 433, 436 or 451 of SEQ ID NO: 2, or anequivalent position in the parent glucoamylase.
 9. The glucoamylasevariant of claim 1, wherein the variant further comprises one or moreamino acid substitutions corresponding to position: 4, 5, 24, 29, 43,44, 49, 70, 75, 76, 100, 108, 119, 124, 137, 146, 148, 169, 171, 172,175, 178, 181, 208, 211, 243, 292, 294, 297, 314, 316, 317, 340, 341,350, 356, 363, 368, 369, 376, 395, 401, 412, 433, 436 or 451 of SEQ IDNO: 2, or an equivalent position in a parent glucoamylase.
 10. Theglucoamylase variant of claim 1, wherein the glucoamylase variantfurther comprises one or more amino acid substitutions corresponding toposition: 5, 24, 43, 44, 49, 70, 75, 76, 94, 119, 141, 146, 148, 172,175, 178, 180, 181, 208, 211, 243, 294, 309, 314, 353, 369, 375, or 409of SEQ ID NO: 2, or an equivalent position in a parent glucoamylase. 11.The glucoamylase variant of claim 1, wherein the glucoamylase variantfurther comprises one or more amino acid substitutions corresponding toposition: 43, 44, or 294 of SEQ ID NO: 2, or an equivalent position in aparent glucoamylase.
 12. The glucoamylase variant of claim 1, whereinthe parent glucoamylase is selected from a glucoamylase obtained from aTrichoderma spp., an Aspergillus spp., a Humicola spp., a Penicilliumspp., a Talaromyces spp., or a Schizosaccharomyces spp.
 13. Theglucoamylase variant of claim 12, wherein the parent glucoamylase isobtained from a Trichoderma spp. or an Aspergillus spp.
 14. Theglucoamylase variant of claim 1, wherein the variant comprises two ormore amino acid substitutions in a residue corresponding to position:N61I, G73F, L417R/V, T430A/M, A431L/Q, E503A/V, Q511H, A535R, A539R, orN563I/K of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase.
 15. The glucoamylase variant of claim 14, wherein thevariant further comprises one or more of the following substitutions:D4L/E/R/S/C/A/Q/W, F5C/M/N/R/S/T/V/W, I12L/R, D24E/L/Y/T,F29L/I/D/C/S/V/W, I43F/R/D/Y/S/Q, D44E/H/K/S/N/Y/F/R/C, Y47W, Y49N,Q70R/K/M/P/G/L/F, Q75R/K/A, R76L/M/K/T/P, P94L, D100W/I/Q/M/P/A/N,N119P/T/Y/D/E, N146S/G/C/H/E/D/T/W/L/F/M, Q148V/Y/H/A/C/D/G/M/R/S/T,Y169D/F, Q172C/A/D/R/E/F/H/V/L/M/N/S/T/V, F175H/A/G/R/S/T/C/W/Y,W178A/C/D/E/F/G/H/K/N/R/S/T/V/Y, E180A/C/G/H/I/L/N/P/Q/R/S/T/V/Y/,V181E/C/D/G/H/I/P/T/Y/S/L/K/F/A, Q208L/A/C/E/N/F/H/T,S211C/R/E/A/Y/W/M/H/L/I/R/Q/T, E243 S/R/N/M/Y/A/L, R245A/E/M/I/P/V,I292D/H/P/R/T/N/V/F/L, G294C/D/E/T/Q/I/A,K297F/L/P/T/M/D/N/Q/A/Y/H/S/R/W, R309A/C/G/H/I/N/P/Q/S/T/W/Y/L,Y310E/G/L/P/S/W/R/Q, D313Q, V314A/R/N/D/C/E/Q/G/H/I/L/K/M/F/P/S/T/W/Y,Y315F, Y316Q/R, N317T/H, K340D/T, K341F/D/P/V/G/S, T350S/E/A/N,Q356H/D/E, T363L/R/C/H/W, S368W/D/F/L, S369F, N376Q/T/H/S/V, Y395Q/R/S,A398S/I/T, S401C/v, R408S, N409W/T/K, T412A/H/K/G, R433H/Q, I436A/T, orS451M/T/H of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase.
 16. The glucoamylase variant of claim 14, wherein thevariant further comprises one or more of the following substitutions:I43F/R/D/Y/S/Q, D44E/H/K/S/N/Y/F/R/C, or G294C/D/E/T/Q/I/A of SEQ ID NO:2, or an equivalent position in a parent glucoamylase.
 17. Theglucoamylase variant of claim 1, wherein the glucoamylase variantexhibits altered thermostability as compared to the parent glucoamylase.18. The glucoamylase variant of claim 17, wherein the glucoamylasevariant exhibits increased thermostability as compared to the parentglucoamylase.
 19. The glucoamylase variant of claim 1, wherein theglucoamylase variant exhibits altered specific activity as compared tothe parent glucoamylase.
 20. The glucoamylase variant of claim 19,wherein the glucoamylase variant exhibits increased specific activity ascompared to the parent glucoamylase.
 21. The glucoamylase variant ofclaim 1, wherein the glucoamylase variant exhibits both increasedthermostability and increased specific activity as compared to theparent glucoamylase.
 22. A polynucleotide encoding the variant ofclaim
 1. 23. A vector comprising the polynucleotide of claim
 22. 24. Ahost cell comprising the vector of claim
 23. 25. An enzyme compositioncomprising the glucoamylase variant of claim
 1. 26. The enzymecomposition of claim 25 wherein the enzyme composition is used in astarch conversion process.
 27. The enzyme composition of claim 25,wherein the enzyme composition is used in an alcohol fermentationprocess.
 28. A method of producing a glucoamylase variant in a host cellcomprising culturing the host cell of claim 24 under conditions suitablefor the expression and production of the glucoamylase variant andproducing the glucoamylase variant.
 29. The method according to claim 28further comprising recovering the glucoamylase variant from the culture.30. A glucoamylase variant comprising amino acid substitutions atresidues corresponding to positions: I43Q/D44C, D44C/G294C, I43Q/G294C,or I43Q/D44C/G294 of SEQ ID NO: 2, or an equivalent position in a parentglucoamylase, wherein the glucoamylase variant has at least 80% sequenceidentity with SEQ ID NO: 1, 2, 3, 5, 6, 7, 8, or 9.